Isoindolinone inhibitors of phosphatidylinositol 3-kinase

ABSTRACT

The present invention relates to compounds useful as inhibitors of PI3K, particularly of PI3Kγ. The invention also provides pharmaceutically acceptable compositions comprising said compounds and methods of using the compositions in the treatment of various disease, conditions, or disorders.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to compounds useful as inhibitors ofphosphatidylinositol 3-kinase (PI3K). The invention also providespharmaceutically acceptable compositions comprising the compounds of theinvention and methods of using the compositions in the treatment ofvarious disorders.

BACKGROUND OF THE INVENTION

PI3Ks are a family of lipid kinases that catalyze the phosphorylation ofthe membrane lipid phosphatidylinositol (PI) on the 3′-OH of theinositol ring to produce PI 3-phosphate [PI(3)P, PIP], PI3,4-bisphosphate [PI(3,4)P₂, PIP2] and PI 3,4,5-trisphosphate[PI(3,4,5)P₃, PIP3]. PI(3,4)P₂ and PI(3,4,5)P₃ act as recruitment sitesfor various intracellular signaling proteins, which in turn formsignaling complexes to relay extracellular signals to the cytoplasmicface of the plasma membrane.

Eight mammalian PI3Ks have been identified so far, including four classI PI3Ks. Class Ia includes PI3Kα, PI3Kβ and PI3Kδ. All of the class Iaenzymes are heterodimeric complexes comprising a catalytic subunit(p110α, p 110β or p110δ) associated with an SH2 domain-containing p85adapter subunit. Class Ia PI3Ks are activated through tyrosine kinasesignaling and are involved in cell proliferation and survival. PI3Kα andPI3Kβ have also been implicated in tumorigenesis in a variety of humancancers. Thus, pharmacological inhibitors of PI3Kα and PI3Kβ are usefulfor treating various types of cancer.

PI3Kγ, the only member of the Class Ib PI3Ks, consists of a catalyticsubunit p110γ, which is associated with a p101 regulatory subunit. PI3Kγis regulated by G protein-coupled receptors (GPCRs) via association withβγ subunits of heterotrimeric G proteins. PI3Kγ is expressed primarilyin hematopoietic cells and cardiomyocytes and is involved ininflammation and mast cell function. Thus, pharmacological inhibitors ofPI3Kγ are useful for treating a variety of inflammatory diseases,allergies and cardiovascular diseases.

Although a number of PI3K inhibitors have been developed, there is aneed for additional compounds to inhibit PI3Ks for treating variousdisorders and diseases, especially those affecting the central nervoussystem (CNS). Accordingly, it would be desirable to develop additionalcompounds that are useful as inhibitors of PI3K that penetrate theblood-brain barrier (BBB).

SUMMARY OF THE INVENTION

It has been found that compounds of this invention, and pharmaceuticallyacceptable compositions thereof, are effective as inhibitors of PI3K,particularly PI3Kγ. Accordingly, the invention features compounds havingthe general formula:

or a pharmaceutically acceptable salt thereof, where each of A, B, C, D,E, X¹, X², R¹, R², R³, and R⁴ is as defined herein.

The invention also provides pharmaceutical compositions that include acompound of formula I and a pharmaceutically acceptable carrier,adjuvant, or vehicle. These compounds and pharmaceutical compositionsare useful for treating or lessening the severity of a variety ofdisorders, including autoimmune diseases and inflammatory diseases ofthe CNS.

The compounds and compositions provided by this invention are alsouseful for the study of PI3K in biological and pathological phenomena;the study of intracellular signal transduction pathways mediated by suchkinases; and the comparative evaluation of new kinase inhibitors.

DETAILED DESCRIPTION OF THE INVENTION Definitions and GeneralTerminology

As used herein, the following definitions shall apply unless otherwiseindicated. For purposes of this invention, the chemical elements areidentified in accordance with the Periodic Table of the Elements, CASversion, and the Handbook of Chemistry and Physics, 75^(th) Ed. 1994.Additionally, general principles of organic chemistry are described in“Organic Chemistry,” Thomas Sorrell, University Science Books,Sausalito: 1999, and “March's Advanced Organic Chemistry,” 5^(th) Ed.,Smith, M. B. and March, J., eds. John Wiley & Sons, New York: 2001, theentire contents of which are hereby incorporated by reference.

Compounds that have been drawn with stereochemical centers defined arestereochemically pure, but with the absolute stereochemistry stillundefined. Such compounds can have either the R or S configuration. Inthose cases where such an absolute assignment has been determined, thechiral center(s) will be labeled R or S in the drawing.

As described herein, compounds of the invention may optionally besubstituted with one or more substituents, such as are illustratedgenerally above, or as exemplified by particular classes, subclasses,and species of the invention. It will be appreciated that the phrase“optionally substituted” is used interchangeably with the phrase“substituted or unsubstituted.” In general, the term “substituted,”whether preceded by the term “optionally” or not, refers to thereplacement of one or more hydrogen radicals in a given structure withthe radical of a specified substituent. Unless otherwise indicated, anoptionally substituted group may have a substituent at eachsubstitutable position of the group. When more than one position in agiven structure can be substituted with more than one substituentselected from a specified group, the substituent may be either the sameor different at each position.

As described herein, when the term “optionally substituted” precedes alist, said term refers to all of the subsequent substitutable groups inthat list. For example, if X is halogen; optionally substituted C₁₋₃alkyl or phenyl; X may be either optionally substituted alkyl oroptionally substituted phenyl. Likewise, if the term “optionallysubstituted” follows a list, said term also refers to all of thesubstitutable groups in the prior list unless otherwise indicated. Forexample: if X is halogen, C₁₋₃ alkyl, or phenyl, wherein X is optionallysubstituted by J^(X), then both C₁₋₃ alkyl and phenyl may be optionallysubstituted by J^(X). As is apparent to one having ordinary skill in theart, groups such as H, halogen, NO₂, CN, NH₂, OH, or OCF₃ would not beincluded because they are not substitutable groups. If a substituentradical or structure is not identified or defined as “optionallysubstituted,” the substituent radical or structure is unsubstituted.

Combinations of substituents envisioned by this invention are preferablythose that result in the formation of stable or chemically feasiblecompounds. The term “stable,” as used herein, refers to compounds thatare not substantially altered when subjected to conditions to allow fortheir production, detection, and, preferably, their recovery,purification, and use for one or more of the purposes disclosed herein.In some embodiments, a stable compound or chemically feasible compoundis one that is not substantially altered when kept at a temperature of40° C. or less, in the absence of moisture or other chemically reactiveconditions, for at least a week.

The term “aliphatic” or “aliphatic group,” as used herein, means astraight-chain (i.e., unbranched) or branched, substituted orunsubstituted hydrocarbon chain that is completely saturated or thatcontains one or more units of unsaturation. Unless otherwise specified,aliphatic groups contain 1-20 carbon atoms. In some embodiments,aliphatic groups contain 1-10 carbon atoms. In other embodiments,aliphatic groups contain 1-8 carbon atoms. In still other embodiments,aliphatic groups contain 1-6 carbon atoms, and in yet other embodiments,aliphatic groups contain 1-4 carbon atoms. Suitable aliphatic groupsinclude, but are not limited to, linear or branched, substituted orunsubstituted alkyl, alkenyl, or alkynyl groups. Further examples ofaliphatic groups include methyl, ethyl, propyl, butyl, isopropyl,isobutyl, vinyl, and sec-butyl. The terms “alkyl” and the prefix “alk-,”as used herein, are inclusive of both straight chain and branchedsaturated carbon chain. The term “alkylene,” as used herein, representsa saturated divalent straight or branched chain hydrocarbon group and isexemplified by methylene, ethylene, isopropylene and the like. The term“alkylidene,” as used herein, represents a divalent straight chain alkyllinking group. The term “alkenyl,” as used herein, represents monovalentstraight or branched chain hydrocarbon group containing one or morecarbon-carbon double bonds. The term “alkynyl,” as used herein,represents a monovalent straight or branched chain hydrocarbon groupcontaining one or more carbon-carbon triple bonds.

The term “cycloaliphatic” (or “carbocycle”) refers to a monocyclic C₃-C₈hydrocarbon or bicyclic C₈-C₁₂ hydrocarbon that is completely saturatedor that contains one or more units of unsaturation, but which is notaromatic, that has a single point of attachment to the rest of themolecule, and wherein any individual ring in said bicyclic ring systemhas 3-7 members. Suitable cycloaliphatic groups include, but are notlimited to, cycloalkyl, cycloalkenyl, and cycloalkynyl. Further examplesof aliphatic groups include cyclopentyl, cyclopentenyl, cyclohexyl,cyclohexenyl, cycloheptyl, and cycloheptenyl.

The term “heterocycle,” “heterocyclyl,” “heterocycloaliphatic,” or“heterocyclic” as used herein refers to a monocyclic, bicyclic, ortricyclic ring system in which at least one ring in the system containsone or more heteroatoms, which is the same or different, and that iscompletely saturated or that contains one or more units of unsaturation,but which is not aromatic, and that has a single point of attachment tothe rest of the molecule. In some embodiments, the “heterocycle,”“heterocyclyl,” “heterocycloaliphatic,” or “heterocyclic” group hasthree to fourteen ring members in which one or more ring members is aheteroatom independently selected from oxygen, sulfur, nitrogen, orphosphorus, and each ring in the system contains 3 to 8 ring members.

Examples of heterocyclic rings include, but are not limited to, thefollowing monocycles: 2-tetrahydrofuranyl, 3-tetrahydrofuranyl,2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl, 2-morpholino,3-morpholino, 4-morpholino, 2-thiomorpholino, 3-thiomorpholino,4-thiomorpholino, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl,1-tetrahydropiperazinyl, 2-tetrahydropiperazinyl,3-tetrahydropiperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl,1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl,1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl,2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl,2-imidazolidinyl, 4-imidazolidinyl, 5-imidazolidinyl; and the followingbicycles: 3-1H-benzimidazol-2-one, 3-(1-alkyl)-benzimidazol-2-one,indolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane,benzodithiane, and 1,3-dihydro-imidazol-2-one.

The term “heteroatom” means one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon, including any oxidized form of nitrogen, sulfur,or phosphorus; the quaternized form of any basic nitrogen; or asubstitutable nitrogen of a heterocyclic ring, for example N (as in3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR⁺ (as inN-substituted pyrrolidinyl).

The term “unsaturated,” as used herein, means that a moiety has one ormore units of unsaturation.

The term “alkoxy,” or “thioalkyl,” as used herein, refers to an alkylgroup, as previously defined, attached to the principal carbon chainthrough an oxygen (“alkoxy”) or sulfur (“thioalkyl”) atom.

The terms “haloalkyl,” “haloalkenyl,” and “haloalkoxy” mean alkyl,alkenyl, or alkoxy, as the case may be, substituted with one or morehalogen atoms. The term “halogen” means F, Cl, Br, or I.

The term “aryl” used alone or as part of a larger moiety as in“aralkyl,” “aralkoxy,” or “aryloxyalkyl,” refers to a monocyclic,bicyclic, or tricyclic carbocyclic ring system having a total of six tofourteen ring members, wherein said ring system has a single point ofattachment to the rest of the molecule, at least one ring in the systemis aromatic and wherein each ring in the system contains 3 to 7 ringmembers. The term “aryl” may be used interchangeably with the term “arylring.” Examples of aryl rings include phenyl, naphthyl, and anthracene.

The term “heteroaryl,” used alone or as part of a larger moiety as in“heteroaralkyl,” or “heteroarylalkoxy,” refers to a monocyclic,bicyclic, and tricyclic ring system having a total of five to fourteenring members, wherein said ring system has a single point of attachmentto the rest of the molecule, at least one ring in the system isaromatic, at least one ring in the system contains one or moreheteroatoms independently selected from nitrogen, oxygen, sulfur orphosphorus, and wherein each ring in the system contains 3 to 7 ringmembers. The term “heteroaryl” may be used interchangeably with the term“heteroaryl ring” or the term “heteroaromatic.”

Further examples of heteroaryl rings include the following monocycles:2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl,5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl,4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl,3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl,pyridazinyl (e.g., 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl,5-thiazolyl, tetrazolyl (e.g., 5-tetrazolyl), triazolyl (e.g.,2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, pyrazolyl (e.g.,2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl,1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl,1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, pyrazinyl, 1,3,5-triazinyl, andthe following bicycles: benzimidazolyl, benzofuryl, benzothiophenyl,indolyl (e.g., 2-indolyl), purinyl, quinolinyl (e.g., 2-quinolinyl,3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl,3-isoquinolinyl, or 4-isoquinolinyl).

In some embodiments, an aryl (including aralkyl, aralkoxy, aryloxyalkyl,and the like) or heteroaryl (including heteroaralkyl, heteroarylalkoxy,and the like) group may contain one or more substituents. Suitablesubstituents on the unsaturated carbon atom of an aryl or heteroarylgroup include: halogen; C₁₋₄aliphatic, —OH; —OR°; —SH°; —SR°;1,2-methylenedioxy; 1,2-ethylenedioxy; phenyl (Ph); —O(Ph);—(CH₂)₁₋₂(Ph); —CH═CH(Ph); —NO₂; —CN; —NH₂; —NH(R°); N(R°)₂; —NHC(O)R°;—NR°C(O)R°; —NHC(S)R°; —NR°C(S)R°; —NHC(O)NH₂; —NHC(O)NH(R°);—NHC(O)N(R°)₂; —NR°C(O)NH(R°; —NR°C(O)N(R°)₂; —NHC(S)NH₂; —NHC(S)N(R°)₂;—NHC(S)NH(R°); —NR°C(S)NH(R°); —NR°C(S)N(R°)₂; —NHC(O)OR°; —NR°C(O)OR°;—C(O)OH; —C(O)OR°; —C(O)R°; —C(S)R°; —C(O)NH₂; —C(O)NH(R°); —C(O)N(R°)₂;—C(S)NH₂; —C(S)NH(R°); —C(S)N(R°)₂; —OC(O)NH₂; —OC(O)NH(R°);—OC(O)N(R°)₂; —OC(O)R°); —C(NOR°H; —C(NOR°)R°; —S(O)₂R°; —S(O)₃R°;—S(O)₃H; —S(O)₂NH₂; —S(O)₂NH(R°); —S(O)₂N(R°)₂; —S(O)R°; —NHS(O)₂R°;—NR°S(O)₂R°; —N(OR°)R°; —(CH₂)₀₋₂NHC(O)R°; -L-R°; -L-N(R°)₂; -L-SR°;-L-OR°; -L-(C₃₋₁₀ cycloaliphatic), -L-(C₆₋₁₀ aryl), -L-(5-10 memberedheteroaryl), -L-(5-10 membered heterocyclyl), oxo, C₁₋₄ haloalkoxy, C₁₋₄haloalkyl, -L-NO₂, -L-CN, -L-OH, -L-CF₃; or two substituents, on thesame carbon or on different carbons, together with the carbon orintervening carbons to which they are bound, form a 5-7 memberedsaturated, unsaturated, or partially saturated ring, wherein L is a C₁₋₆alkylene group in which up to three methylene units are replaced by—NH—, —NR°—, —O—, —S—, —C(O)O—, —OC(O)—, —C(O)CO—, —C(O)—, —C(O)NH—,—C(O)NR°—, —C(═N—CN), —NHCO—, —NR°CO—, —NHC(O)O—, —NR°C(O)O—, —S(O)₂NH—,—S(O)₂NR°—, —NHS(O)₂—, —NR°S(O)₂—, —NHC(O)NH—, —NR°C(O)NH—, —NHC(O)NR°—,—NR°C(O)NR°, —OC(O)NH—, —OC(O)NR°—, —NHS(O)₂NH—, —NR°S(O)₂NH—,—NHS(O)₂NR°—, —NR°S(O)₂NR°—, —S(O)—, or —S(O)₂—, and wherein eachoccurrence of R° is independently selected from optionally substitutedC₁₋₆ aliphatic, an unsubstituted 5 to 6 membered heteroaryl orheterocyclic ring, phenyl, or —CH₂(Ph), or, two independent occurrencesof R°, on the same substituent or different substituents, taken togetherwith the atom(s) to which each R° group is bound, form a 5-8-memberedheterocyclyl, aryl, or heteroaryl ring or a 3- to 8-membered cycloalkylring, wherein said heteroaryl or heterocyclyl ring has 1 to 3heteroatoms independently selected from nitrogen, oxygen, or sulfur.Non-limiting optional substituents on the aliphatic group of R° include—NH₂, —NH(C₁₋₄ aliphatic), —N(C₁₋₄ aliphatic)₂, halogen, C₁₋₄ aliphatic,—OH, —O(C₁₋₄ aliphatic), —NO₂, —CN, —C(O)OH, —C(O)O(C₁₋₄ aliphatic),—O(haloC₁₋₄ aliphatic), or haloC₁₋₄ aliphatic, wherein each of theforegoing C₁₋₄ aliphatic groups of R° is unsubstituted.

In some embodiments, an aliphatic or heteroaliphatic group, or anon-aromatic heterocyclic ring may contain one or more substituents.Suitable substituents on the saturated carbon of an aliphatic orheteroaliphatic group, or of a non-aromatic heterocyclic ring areselected from those listed above for the unsaturated carbon of an arylor heteroaryl group and additionally include the following: ═O, ═S,═NNHR*, ═NN(R*)₂, ═NNHC(O)R*, ═NNHC(O)O(alkyl), ═NNHS(O)₂(alkyl), or═NR*, where each R* is independently selected from hydrogen or anoptionally substituted C₁₋₈ aliphatic. Optional substituents on thealiphatic group of R* are selected from —NH₂, —NH(C₁₋₄ aliphatic),—N(C₁₋₄ aliphatic)₂, halogen, C₁₋₄ aliphatic, —OH, —O(C₁₋₄ aliphatic),—NO₂, —CN, —C(O)OH, —C(O)O(C₁₋₄ aliphatic), —C(O)NH₂, —C(O)NH(C₁₋₄aliphatic), —C(O)N(C₁₋₄ aliphatic)₂, —O(halo-C₁₋₄ aliphatic), andhalo(C₁₋₄ aliphatic), where each of the foregoing C₁₋₄ aliphatic groupsof R* is unsubstituted; or two R* on the same nitrogen are takentogether with the nitrogen to form a 5-8 membered heterocyclyl orheteroaryl ring having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur.

In some embodiments, optional substituents on the nitrogen of anon-aromatic heterocyclic ring include —R⁺, —N(R⁺)₂, —C(O)R⁺, —C(O)OR⁺,—C(O)C(O)R⁺, —C(O)CH₂C(O)R⁺, —S(O)₂R⁺, —S(O)₂N(R⁺)₂, —C(═S)N(R⁺)₂,—C(═NH)—N(R⁺)₂, or —NR⁺S(O)₂R⁺; wherein R⁺ is hydrogen, an optionallysubstituted C₁₋₆ aliphatic, optionally substituted phenyl, optionallysubstituted —O(Ph), optionally substituted —CH₂(Ph), optionallysubstituted —(CH₂)₁₋₂(Ph); optionally substituted —CH═CH(Ph); or anunsubstituted 5-6 membered heteroaryl or heterocyclic ring having one tofour heteroatoms independently selected from oxygen, nitrogen, orsulfur, or, two independent occurrences of R⁺, on the same substituentor different substituents, taken together with the atom(s) to which eachR⁺ group is bound, form a 5-8-membered heterocyclyl, aryl, or heteroarylring or a 3-8 membered cycloalkyl ring, wherein said heteroaryl orheterocyclyl ring has 1-3 heteroatoms independently selected fromnitrogen, oxygen, or sulfur. Optional substituents on the aliphaticgroup or the phenyl ring of R⁺ are selected from —NH₂, —NH(C₁₋₄aliphatic), —N(C₁₋₄ aliphatic)₂, halogen, C₁₋₄ aliphatic, —OH, —O(C₁₋₄aliphatic), —NO₂, —CN, —C(O)OH, —C(O)O(C₁₋₄ aliphatic), —O(halo(C₁₋₄aliphatic)), or halo(C₁₋₄ aliphatic), wherein each of the foregoingC₁₋₄aliphatic groups of R⁺ is unsubstituted.

As detailed above, in some embodiments, two independent occurrences ofR° (or R⁺, or any other variable similarly defined herein), may be takentogether with the atom(s) to which each variable is bound to form a5-8-membered heterocyclyl, aryl, or heteroaryl ring or a 3-8-memberedcycloalkyl ring. Exemplary rings that are formed when two independentoccurrences of R° (or R⁺, or any other variable similarly definedherein) are taken together with the atom(s) to which each variable isbound include, but are not limited to the following: a) two independentoccurrences of R° (or R⁺, or any other variable similarly definedherein) that are bound to the same atom and are taken together with thatatom to form a ring, for example, N(R°)₂, where both occurrences of R°are taken together with the nitrogen atom to form a piperidin-1-yl,piperazin-1-yl, or morpholin-4-yl group; and b) two independentoccurrences of R° (or R⁺, or any other variable similarly definedherein) that are bound to different atoms and are taken together withboth of those atoms to form a ring, for example where a phenyl group issubstituted with two occurrences of

these two occurrences of R° are taken together with the oxygen atoms towhich they are bound to form a fused 6-membered oxygen containing ring:

It will be appreciated that a variety of other rings can be formed whentwo independent occurrences of R° (or R⁺, or any other variablesimilarly defined herein) are taken together with the atom(s) to whicheach variable is bound and that the examples detailed above are notintended to be limiting.

In some embodiments, a methylene unit of the alkyl or aliphatic chain isoptionally replaced with another atom or group. Examples of such atomsor groups would include, but are not limited to, —NR°—, —O—, —S—,—C(O)O—, —OC(O)—, —C(O)CO—, —C(O)—, —C(O)NR°—, —C(═N—CN), —NR°CO—,—NR°C(O)O—, —S(O)₂NR°—, —NR°S(O)₂—, —NR°C(O)NR°—, —OC(O)NR°—,—NR°S(O)₂NR°—, —S(O)—, or —S(O)₂—, wherein R° is defined herein. Unlessotherwise specified, the optional replacements form a chemically stablecompound. Optional atom or group replacements can occur both within thechain and at either end of the chain; i.e. both at the point ofattachment and/or also at the terminal end. Two optional replacementscan also be adjacent to each other within a chain so long as it resultsin a chemically stable compound. Unless otherwise specified, if thereplacement occurs at the terminal end, the replacement atom is bound toan H on the terminal end. For example, if one methylene unit of—CH₂CH₂CH₃ was optionally replaced with —O—, the resulting compoundcould be —OCH₂CH₃, —CH₂OCH₃, or —CH₂CH₂OH.

As described herein, a bond drawn from a substituent to the center ofone ring within a multiple-ring system (as shown below) representssubstitution of the substituent at any substitutable position in any ofthe rings within the multiple ring system. For example, Structure arepresents possible substitution in any of the positions shown inStructure b.

This also applies to multiple ring systems fused to optional ringsystems (which would be represented by dotted lines). For example, inStructure c, X is an optional substituent both for ring A and ring B.

If, however, two rings in a multiple ring system each have differentsubstituents drawn from the center of each ring, then, unless otherwisespecified, each substituent only represents substitution on the ring towhich it is attached. For example, in Structure d, Y is an optionallysubstituent for ring A only, and X is an optional substituent for ring Bonly.

The term “protecting group,” as used herein, represent those groupsintended to protect a functional group, such as, for example, analcohol, amine, carboxyl, carbonyl, etc., against undesirable reactionsduring synthetic procedures. Commonly used protecting groups aredisclosed in Greene and Wuts, Protective Groups In Organic Synthesis,3^(rd) Edition (John Wiley & Sons, New York, 1999), which isincorporated herein by reference. Examples of nitrogen protecting groupsinclude acyl, aroyl, or carbamyl groups such as formyl, acetyl,propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl,trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl,α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl,4-nitrobenzoyl and chiral auxiliaries such as protected or unprotectedD, L or D, L-amino acids such as alanine, leucine, phenylalanine and thelike; sulfonyl groups such as benzenesulfonyl, p-toluenesulfonyl and thelike; carbamate groups such as benzyloxycarbonyl,p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl,p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl,p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl,3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl,3,4,5-trimethoxybenzyloxycarbonyl,1-(p-biphenylyl)-1-methylethoxycarbonyl,α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl,t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl,ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl,2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl,adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and thelike, arylalkyl groups such as benzyl, triphenylmethyl, benzyloxymethyland the like and silyl groups such as trimethylsilyl and the like.Preferred N-protecting groups are formyl, acetyl, benzoyl, pivaloyl,t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc)and benzyloxycarbonyl (Cbz).

The term “prodrug,” as used herein, represents a compound that istransformed in vivo into a compound of formula I or a compound listed inTable 1. Such a transformation can be affected, for example, byhydrolysis in blood or enzymatic transformation of the prodrug form tothe parent form in blood or tissue. Prodrugs of the compounds of theinvention may be, for example, esters. Esters that may be utilized asprodrugs in the present invention are phenyl esters, aliphatic (C₁-C₂₄)esters, acyloxymethyl esters, carbonates, carbamates, and amino acidesters. For example, a compound of the invention that contains an OHgroup may be acylated at this position in its prodrug form. Otherprodrug forms include phosphates, such as, for example those phosphatesresulting from the phosphonation of an OH group on the parent compound.A thorough discussion of prodrugs is provided in T. Higuchi and V.Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S.Symposium Series, Edward B. Roche, ed., Bioreversible Carriers in DrugDesign, American Pharmaceutical Association and Pergamon Press, 1987,and Judkins et al., Synthetic Communications 26(23):4351-4367, 1996,each of which is incorporated herein by reference.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, (Z) and (E) double bondisomers, and (Z) and (E) conformational isomers. Therefore, singlestereochemical isomers as well as enantiomeric, diastereomeric, andgeometric (or conformational) mixtures of the present compounds arewithin the scope of the invention.

Unless otherwise stated, all tautomeric forms of the compounds of theinvention are within the scope of the invention. Additionally, unlessotherwise stated, structures depicted herein are also meant to includecompounds that differ only in the presence of one or more isotopicallyenriched atoms. For example, compounds having the present structuresexcept for the replacement of hydrogen by deuterium or tritium, or thereplacement of a carbon by a ¹³C- or ¹⁴C-enriched carbon are within thescope of this invention. Such compounds are useful, for example, asanalytical tools, probes in biological assays, or as PI3K inhibitorswith improved therapeutic profile.

Description of Compounds of the Invention

In one aspect, the invention features compounds having formula I:

-   or a pharmaceutically acceptable salt thereof, wherein:-   X¹ is N or CH;-   X² is N, CH, or C—CH₃;-   R¹ is selected from a phenyl ring, a 5-6 membered heteroaryl ring, a    pyridone ring, or a 9-10 membered fused bicyclic heteroaryl or    heterocyclic ring system wherein each of said rings or ring systems    is optionally substituted with 1 or 2 independent occurrences of    R^(1a) and each of said heteroaryl or heterocyclic rings has 1, 2,    or 3 heteroatoms selected from nitrogen, oxygen, or sulfur;-   R^(1a) is chloro, fluoro, C₁₋₈aliphatic,    —(CH₂)₀₋₂C₃₋₆cycloaliphatic, —(CH₂)₀₋₂-5-6 membered heterocyclic    having up to two heteroatoms selected from nitrogen, oxygen, or    sulfur, —CN, —C(O)C₁₋₄aliphatic, —C(O)NH(C₁₋₄aliphatic),    —C(O)N(C₁₋₄aliphatic)₂, —C(O)OC₁₋₄aliphatic, —S(O)₂NH(C₁₋₄    aliphatic), —S(O)₂N(C₁₋₄ aliphatic)₂, or —S(O)₂C₁₋₄ aliphatic,    wherein up to 3 non-adjacent carbon atoms of said aliphatic or    cycloaliphatic of R^(1a) can be substituted for by —O—, —S—, or    —N(R^(1b))— and wherein each of said aliphatic, cycloaliphatic, or    heterocyclic of R^(1a) is optionally and independently substituted    with up to 4 occurrences of J^(R);-   each J^(R) is independently fluoro, oxo, —(CH₂)₀₋₂CN, —(CH₂)₀₋₂CF₃,    —C(O)R^(1b), —C(O)N(R^(1b))₂, —C(O)O(R^(1b)), —N(R^(1b))₂,    —N(R^(1b))C(O)R^(1b), —(CH₂)₀₋₂OR^(1b), phenyl, or a 5-6 membered    heteroaryl, 4-6 heterocyclyl, or 9-11 fused bicyclic heteroaryl or    heterocyclyl, each of said heteroaryl or heterocyclyl rings having    up to 3 atoms selected from nitrogen, oxygen, or sulfur, wherein    each of said cycloaliphatic, phenyl, heteroaryl, or heterocyclyl is    optionally substituted with up to 2 R^(1c);-   each R^(1b) is, independently, selected from hydrogen,    C₁₋₈aliphatic, —(CH₂)₀₋₁C₃₋₆cycloaliphatic,    —(CH₂)₀₋₁C₄₋₆heterocyclic having up to two heteroatoms selected from    N or O, or two R^(1b) together with the atom to which they are    bonded form a 5-6 membered heterocyclic ring, wherein each    aliphatic, cycloaliphatic, or heterocyclic is optionally substituted    with up to three F atoms or up to two —OH, —C₁₋₂alkyl, or    —OC₁₋₂alkyl groups;-   each R^(1c) is, independently, fluoro, chloro, C₁₋₄aliphatic,    —(CH₂)₀₋₂OH, —CN, —C(O)C₁₋₄aliphatic, or —C(O)OC₁₋₄aliphatic;-   R² is hydrogen, F, Cl, CF₃, C₁₋₂aliphatic, C₃₋₄cycloaliphatic,    —N(CH₃)₂, —N(CH₂)₃, —OCF₃, —OCHF₂, or —OC₁₋₂aliphatic;-   R³ is hydrogen, C₁₋₆aliphatic, C₃₋₆ cycloaliphatic, C₄₋₇    heterocyclyl having 1 or 2 atoms selected from N or O, —(CH₂)₀₋₁CF₃,    —OH, —OC₁₋₆aliphatic, —OC₃₋₆cycloaliphatic, —OC₃₋₆heterocyclyl    having one oxygen atom, —O(CH₂)₂OC₁₋₂aliphatic, or    —OC₁₋₂alkylC(O)OC₁₋₃aliphatic, or benzyl; and-   R⁴ is hydrogen or C₁₋₆alkyl; or R³ and R⁴ together with the carbon    to which they are bonded form a 3-6 membered cycloaliphatic ring, a    3-6 membered heterocyclic ring having up to two atoms selected from    N or O, or a C₂alkenyl, wherein each of said aliphatic,    cycloaliphatic, or heterocyclyl of R³, R⁴, or R³ and R⁴ together is    optionally substituted with up to three F atoms, or up to two    C₁₋₂alkyl, —C(O)C₁₋₄alkyl, —C(O)OC₁₋₄alkyl, —OH, or —OC₁₋₂alkyl    groups;-   A is N or CR^(A);-   B is N or CR^(B), or A=B is a sulfur atom;-   C is N or CR^(C);-   D is N or CR^(D);-   E is N or CR^(E) wherein no more than two of A, B, C, D, or E is N;-   R^(A) is hydrogen, CH₃, or OCH₃;-   R^(B) is hydrogen, F, Cl, C₁₋₃aliphatic, —(CH₂)₀₋₁CF₃,    —(CH₂)₀₋₁CHF₂, or —O(CH₂)₀₋₁CF₃;-   R^(C) is hydrogen, F, Cl, C₁₋₃aliphatic, —(CH₂)₀₋₁CF₃,    —(CH₂)₀₋₁CHF₂, N(R^(1b))₂, —OH, —O(CH₂)₀₋₁CF₃, or —OC₁₋₈aliphatic,    wherein up to 2 non-adjacent carbon atoms of said aliphatic can be    substituted for by —O—;-   R^(D) is hydrogen, fluoro, chloro, C₁₋₄ aliphatic, —C(O)OH,    —C(O)OC₁₋₄ aliphatic, —C(O)N(R^(1b))₂, —CN, —C(R^(D1))═N—OR^(1b),    —N(R^(1b))₂, —N(R^(D1))C(O)C₁₋₄aliphatic, —N(R^(D1))C(O)phenyl,    —N(R^(D1))S(O)₂C₁₋₄aliphatic, —N(R^(D1))S(O)₂N(R^(1b))₂,    —N(R^(D1))S(O)₂phenyl —OH, —OC₁₋₈aliphatic,    —O(CH₂)₀₋₁C₃₋₆cycloaliphatic, —SC₁₋₄aliphatic, —S(O)C₁₋₄aliphatic,    —S(O)₂C₁₋₄aliphatic, or —S(O)₂N(R^(1b))₂; wherein up to 2    non-adjacent carbon atoms of said aliphatic, cycloaliphatic, or    heterocyclic of R^(D) can be substituted for by —O— and each of said    aliphatic, cycloaliphatic, or phenyl of R^(D) can be substituted    with up to 5 fluorine atoms; or R^(D) and R^(C) together with the    atoms to which they are attached form a phenyl or pyridyl ring;-   each R^(D1) is, independently, hydrogen or C₁₋₂alkyl; and-   R^(E) is hydrogen, F, Cl, —NHC(O)C₁₋₈aliphatic, —OH,    —OC₁₋₂aliphatic, —(CH₂)₀₋₁CF₃, —(CH₂)₀₋₁CHF₂, C₁₋₃aliphatic,    C₃₋₄cycloaliphatic, N(R^(1b))₂, azetidin-1-yl.

In one embodiment, compounds have formula I and R^(D) is hydrogen,fluoro, chloro, C₁₋₄aliphatic, —(CH₂)₀₋₁CF₃, —C(O)N(R^(1b))₂, —CN,—N(R^(1b))₂, —NHC(O)C₁₋₈aliphatic, —OH, —O(CH₂)₀₋₁CF₃, —O(CH₂)₀₋₁CHF₂,—O(CH₂)₀₋₁—CH₂F, —OC₁₋₈aliphatic, —O(CH₂)₀₋₁C₃₋₆cycloaliphatic,—SC₁₋₈aliphatic, —S(O)₂C₁₋₈aliphatic, —S(O)₂N(R^(1b))₂; wherein up to 2non-adjacent carbon atoms of said aliphatic, cycloaliphatic, orheterocyclic of R^(D) can be substituted for by —O—, or R^(D) and R^(C)together with the atoms to which they are attached form a phenyl orpyridyl ring; R³ is hydrogen, C₁₋₆alkyl, C₃₋₆ cycloalkyl, —(CH₂)₀₋₁CF₃,—OH, —OC₁₋₆alkyl, —OC₃₋₆cycloalkyl, —OC₃₋₆heterocyclyl having one oxygenatom, —O(CH₂)₂OC₁₋₂alkyl, or —OC₁₋₂alkylC(O)OC₁₋₃alkyl, or benzyl; andR⁴ is hydrogen or C₁₋₆alkyl; or R³ and R⁴ together with the carbon towhich they are bonded form a 3-6 membered cycloalkyl ring, a 3-6membered heterocyclic ring having one oxygen atom, wherein each of saidalkyl, cycloalkyl, or heterocyclyl of R³, R⁴ or R³ and R⁴ together isoptionally substituted with up to two F, C₁₋₂ alkyl or —OC₁₋₂ alkyl.

In another embodiment, compounds have formula I and each R^(1b) is,independently, selected from hydrogen, C₁₋₄ aliphatic, or C₃₋₆cycloaliphatic; R^(B) is hydrogen, F, Cl, —OCF₃, —OC₁₋₂aliphatic, —CF₃,or C₁₋₂aliphatic; R^(C) is hydrogen, F, Cl, C₁₋₃aliphatic, —(CH₂)₀₋₁CF₃,—N(R^(1b))₂, —OH, —OCF₃, or —OC₁₋₈aliphatic; R^(D) is hydrogen, fluoro,chloro, C₁₋₄aliphatic, (CH₂)₀₋₁CF₃, —C(O)NHC₁₋₈aliphatic, —CN,—N(R^(1b))₂, —NHC(O)C₁₋₈aliphatic, —OH, —OCF₃, —OCHF₂, —OC₁₋₈aliphatic,—O(CH₂)₀₋₁C₃₋₆cycloaliphatic, —SC₁₋₈aliphatic, —S(O)₂C₁₋₈aliphatic,—S(O)₂N(R^(1b))₂; wherein up to 2 non-adjacent carbon atoms of saidaliphatic or cycloaliphatic of R^(D) can be substituted for by —O—, orR^(D) and R^(C) together with the atoms to which they are attached forma phenyl or pyridyl ring; R^(E) is hydrogen, F, Cl,—NHC(O)C₁₋₈aliphatic, —OH, —OCF₃, —OC₁₋₂aliphatic, CF₃, C₁₋₂aliphatic,C₃₋₄cycloaliphatic, N(CH₃)₂, azetidin-1-yl; R² is hydrogen, F, Cl, CF₃,C₁₋₂aliphatic, C₃₋₄cycloaliphatic, —N(CH₃)₂, —N(CH₂)₃, —OCF₃, or—OC₁₋₂aliphatic; and R³ is hydrogen, C₁₋₂alkyl, —OH, —OC₁₋₂alkyl,—O(CH₂)₂OC₁₋₂ alkyl, or —OC₁₋₂ alkylC(O)OC₁₋₂ alkyl; and R⁴ is hydrogenor C₁₋₂ alkyl.

In one embodiment, compounds have formula II:

-   or a pharmaceutically acceptable salt thereof, wherein:-   X¹ is CH or N;-   R¹ is selected from a phenyl ring, a 5-membered heteroaryl ring, a    6-membered heteroaryl ring, or a 9- or 10-membered fused bicyclic    heteroaryl or heterocyclic ring system wherein each of said rings or    ring systems is optionally substituted with 1 or 2 independent    occurrences of R^(1a) and each of said heteroaryl or heterocyclic    rings has 1, 2, or 3 heteroatoms selected from nitrogen, oxygen, or    sulfur;-   R^(1a) is chloro, fluoro, C₁₋₆aliphatic, C₃₋₆cycloaliphatic, —CN,    —C(O)R^(1b), —C(O)N(R^(1b))₂, —C(O)O(R^(1b)), or —OR^(1b), wherein    each of said aliphatic or cycloaliphatic is optionally substituted    with up to 3 occurrences of J^(R);-   each J^(R) is independently fluoro, oxo, —CN, —C(O)R^(1b),    —C(O)N(R^(1b))₂, —C(O)O(R^(1b)), —N(R^(1b))₂, —N(R^(1b))C(O)R^(1b),    —OR^(1b), or a 5-membered heteroaryl or heterocyclyl having up to 3    atoms selected from nitrogen, oxygen, or sulfur;-   each R^(1b) is independently selected from hydrogen, C₁₋₄ aliphatic,    or C₃₋₆ cycloaliphatic;-   R² is hydrogen, F, Cl, CF₃, or CH₃;-   B is N;-   C is CR^(C), wherein R^(C) is hydrogen, fluoro, chloro,    C₁₋₃aliphatic, CF₃, —OCF₃, or —OC₁₋₂aliphatic; and-   D is CR^(D), wherein R^(D) is fluoro, chloro, C₁₋₃aliphatic, CF₃,    —OCF₃, or —OC₁₋₂aliphatic.

In one embodiment of compounds of formula II, X¹ is N.

In one embodiment of compounds of formula II, R² is CH₃.

In a further embodiment,

is a substituted pyridine-3-yl.

In another aspect, the invention features compounds having formula I-A:

-   or a pharmaceutically acceptable salt thereof, wherein-   R¹ is

-   wherein-   R^(1a) is —C₁₋₄alkyl, optionally and independently substituted with    —CN, up to three F atoms, or up to two CH₃, —OC₁₋₂alkyl, or —OH    groups;-   R² is C₁₋₂alkyl;-   R³ is hydrogen, —OH, —OC₁₋₄ alkyl, or C₁₋₄alkyl optionally    substituted with up to two —OH groups;-   R⁴ is hydrogen or CH₃, or R³ and R⁴ together form a C₃₋₆cycloalkyl    ring optionally substituted with up to two OH groups, or a 4-6    membered heterocyclic ring having one oxygen or anitrogen atom    optionally substituted with C₁₋₄alkyl, —C(O)C₁₋₄alkyl, or C(O)O    C₁₋₄alkyl;-   R^(C) is hydrogen, F, C₁₋₂alkyl, or —OC₁₋₂alkyl; and-   R^(D) is —OR^(D1), —C(O)N(R^(D1))R^(D2), —S(O)₂N(R^(D1))R^(D2),    —S(O)₁₋₂R^(D2), —N(R^(D1))S(O)₂R^(D2), or    —N(R^(D1))S(O)₂N(R^(D1))R^(D2), wherein-   R^(D1) is hydrogen or C₁₋₂alkyl, and R^(D2) is C₁₋₄alkyl,    —(CH₂)₀₋₁C₃₋₆cycloalkyl, or —(CH₂)₀₋₁C₄₋₆heterocyclyl having up to    two oxygen or nitrogen atoms, each alkyl, cycloalkyl, or    heterocyclyl optionally substituted with up to three F atoms or up    to two —OH groups.

In one embodiment, R^(1a) is C₁₋₂alkyl, optionally substituted with upto 3 fluorine atoms.

In another embodiment, R^(1a) is C₁₋₄alkyl, optionally substituted withCN.

In another embodiment, R² is CH₃.

In another embodiment, at least one of R³ and R⁴ is not hydrogen.

In a further embodiment, each of R³ and R⁴ is CH₃.

In another further embodiment, R³ and R⁴ together form a 4-6 memberedheterocyclic ring having one oxygen or a nitrogen atom optionallysubstituted with C₁₋₄alkyl, —C(O)C₁₋₄alkyl, or —C(O)OC₁₋₄alkyl.

In another embodiment for any of the compounds of formulae I, II, orI-A, R¹ is a 5-membered heteroaryl ring having 1-3 heteroatoms selectedfrom N, O, or S and optionally substituted with 1 or 2 R^(1a) groups.Examples include optionally substituted pyrazol-4-yl, pyrazol-3-yl,imidazol-4-yl, 1,2,3-triazol-4-yl, 1,2,4-triazol-3-yl,1,2,5-triazol-3-yl, 1,3-thiazol-4-yl, 1,3-thiazol-2-yl,1,2-thiazol-5-yl, 1,2-isoxazol-3-yl ring systems.

In another embodiment R¹ is selected from

In another embodiment, R¹ is

In yet another embodiment, R¹ is

In another embodiment, R¹ is selected from

In yet another embodiment, R¹ is

In one embodiment for any of the compounds of formulae I, II, or I-A,

R¹ is

R² is CH₃;

R³ is hydrogen, C₁₋₂alkyl, OH, or OCH₃;R⁴ is hydrogen or CH₃;R^(C) is hydrogen; and

R^(D) is —OC₁₋₂alkyl or —OC₃₋₅cycloalkyl, each optionally substitutedwith up to 3 fluorine atoms.

In a further embodiment, R¹ is 1-(2,2-difluoroethyl)-1H-pyrazol-4-yl or1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl.

In one embodiment for any of the compounds of formulae I, II, or I-A, R¹is a 6-membered heteroaryl ring having 1-3 nitrogens and optionallysubstituted with 1 or 2 R^(1a) groups. In a further embodiment, R¹ is anoptionally substituted pyridyl ring.

In a further embodiment,

R¹ is

R² is CH₃;

R³ is hydrogen, C₁₋₂alkyl, OH, or OCH₃;R⁴ is hydrogen or CH₃;R^(C) is hydrogen, F, Cl, C₁₋₃aliphatic, (CH₂)₀₋₁CF₃, —OCF₃, or—OC₁₋₈aliphatic; andR^(D) is —C(O)NHC₁₋₈aliphatic.

In a another embodiment, R¹ is selected from

In one embodiment for any of the compounds of formulae I, II, or I-A,R^(D) is —C(O)OH, —C(O)N(R^(1b))₂, —CN, —S(O)₂C₁₋₈aliphatic, or—S(O)₂N(R^(1b))₂.

In another embodiment, each of R^(C) and R^(D) is, independently,hydrogen, fluoro, chloro, C₁₋₃aliphatic, CF₃, —OCF₃, —OCHF₂, or—OC₁₋₂aliphatic, wherein at least one of R^(C) and R^(D) is nothydrogen.

In another embodiment, R^(C) is hydrogen and R^(D) is —OC₁₋₃alkyl,optionally substituted with up to 3 F atoms. In a further embodiment,R^(C) is hydrogen and R^(D) is —OCH₃, —OCH₂CH₃, —OCF₃, —OCH₂CF₃, OCHF₂,or OCH₂CHF₂.

In another embodiment, each of R^(C) and R^(D) is —OCH₃.

In one embodiment,

is selected from:

In another embodiment R³ and R⁴, together with the intervening carbonatom, form a 4-6 membered heterocyclic ring having one atom selectedfrom N or O.

In another embodiment, the invention features a compound selected fromthe group of compounds listed in Table 1.

The invention also features a pharmaceutical composition comprising acompound of the invention and a pharmaceutically acceptable carrier,adjuvant, or vehicle.

In one embodiment, the composition includes a therapeutic agent selectedfrom an agent for treating multiple sclerosis, an anti-inflammatoryagent, an immunomodulatory agent, or an immunosuppressive agent.Examples of such additional therapeutic agents include beta interferon,glatiramir, natalizumab, or mitoxantrone.

In another embodiment, the invention features a method of inhibitingPI3K kinase activity in a patient by administering to the patient acompound of formula I, II, or I-A, or a pharmaceutical compositionthereof. In a further embodiment PI3K-gamma is selectively inhibitedover PI3K-alpha, PI3K-beta, or PI3K-gamma. In a further embodiment,PI3K-gamma is selectively inhibited over PI3K-alpha, PI3K-beta, andPI3K-gamma

In another embodiment, the invention features a method of treating orlessening the severity of a disease or condition selected from anautoimmune disease or an inflammatory disease of the brain or spinalcord selected from multiple sclerosis, epilepsy, Parkinson's Disease,Alzheimer's Disease, Huntington's Disease, or amyotrophic lateralsclerosis in a patient by administering to the patient a compound offormula I, II, or I-A, or a pharmaceutical composition thereof.

In a further embodiment, the disease or disorder is multiple sclerosis.

In another embodiment, the method of treatment includes administering toa patient a compound or composition of the invention and an additionaltherapeutic agent, wherein the additional therapeutic agent isappropriate for the disease being treated and is administered togetherwith the compound or composition as a single dosage form, or separatelyas part of a multiple dosage form. Examples of such additionaltherapeutic agents are those useful for treating multiple sclerosis,such as beta interferon, glatiramir, natalizumab, or mitoxantrone.

The invention also features a non-therapeutic method of inhibitingPI3K-gamma kinase activity in a biological sample in vitro comprisingcontacting said biological sample with a compound of formulae I, II, orI-A, or a composition containing said compound.

Compositions, Formulations, and Administration of Compounds of theInvention

In another embodiment, the invention provides a pharmaceuticalcomposition comprising a compound of any of the formulae or classesdescribed herein. In a further embodiment, the invention provides apharmaceutical composition comprising a compound of Table 1. In afurther embodiment, the composition additionally comprises an additionaltherapeutic agent.

According to another embodiment, the invention provides a compositioncomprising a compound of this invention or a pharmaceutically acceptablederivative thereof and a pharmaceutically acceptable carrier, adjuvant,or vehicle. In one embodiment, the amount of compound in a compositionof this invention is such that is effective to measurably inhibit aPI3K, particularly PI3Kγ, in a biological sample or in a patient. Inanother embodiment, the amount of compound in the compositions of thisinvention is such that is effective to measurably inhibit PI3Kα. In oneembodiment, the composition of this invention is formulated foradministration to a patient in need of such composition. In a furtherembodiment, the composition of this invention is formulated for oraladministration to a patient.

The term “patient,” as used herein, means an animal, preferably amammal, and most preferably a human.

It will also be appreciated that certain of the compounds of presentinvention can exist in free form for treatment, or where appropriate, asa pharmaceutically acceptable derivative thereof. According to thepresent invention, a pharmaceutically acceptable derivative includes,but is not limited to, pharmaceutically acceptable prodrugs, salts,esters, salts of such esters, or any other adduct or derivative whichupon administration to a patient in need is capable of providing,directly or indirectly, a compound as otherwise described herein, or ametabolite or residue thereof. As used herein, the term “inhibitoryactive metabolite or residue thereof” means that a metabolite or residuethereof is also an inhibitor of PI3K.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge et al., describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 66:1-19, 1977, which isincorporated herein by reference. Pharmaceutically acceptable salts ofthe compounds of this invention include those derived from suitableinorganic and organic acids and bases. Examples of pharmaceuticallyacceptable, nontoxic acid addition salts are salts of an amino groupformed with inorganic acids such as hydrochloric acid, hydrobromic acid,phosphoric acid, sulfuric acid and perchloric acid or with organic acidssuch as acetic acid, oxalic acid, maleic acid, tartaric acid, citricacid, succinic acid or malonic acid or by using other methods used inthe art such as ion exchange. Other pharmaceutically acceptable saltsinclude adipate, alginate, ascorbate, aspartate, benzenesulfonate,benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate,citrate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate,gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄ alkyl)₄ salts. This inventionalso envisions the quaternization of any basic nitrogen-containinggroups of the compounds disclosed herein. Water or oil-soluble ordispersable products may be obtained by such quaternization.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, C₁₋₈ sulfonate and aryl sulfonate.

As described above, the pharmaceutically acceptable compositions of thepresent invention additionally comprise a pharmaceutically acceptablecarrier, adjuvant, or vehicle, which, as used herein, includes any andall solvents, diluents, or other liquid vehicle, dispersion orsuspension aids, surface active agents, isotonic agents, thickening oremulsifying agents, preservatives, solid binders, lubricants and thelike, as suited to the particular dosage form desired. In Remington: TheScience and Practice of Pharmacy, 21st edition, 2005, ed. D. B. Troy,Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia ofPharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan,1988-1999, Marcel Dekker, New York, the contents of each of which isincorporated by reference herein, are disclosed various carriers used informulating pharmaceutically acceptable compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention.

Some examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, ion exchangers,alumina, aluminum stearate, lecithin, serum proteins, such as humanserum albumin, buffer substances such as phosphates, glycine, sorbicacid, or potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, wool fat, sugars such aslactose, glucose and sucrose; starches such as corn starch and potatostarch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; powdered tragacanth;malt; gelatin; talc; excipients such as cocoa butter and suppositorywaxes; oils such as peanut oil, cottonseed oil; safflower oil; sesameoil; olive oil; corn oil and soybean oil; glycols; such a propyleneglycol or polyethylene glycol; esters such as ethyl oleate and ethyllaurate; agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

The compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal, intraocular,intrahepatic, intralesional, epidural, intraspinal, and intracranialinjection or infusion techniques. Preferably, the compositions areadministered orally, intraperitoneally or intravenously. Sterileinjectable forms of the compositions of this invention may be aqueous oroleaginous suspension. These suspensions may be formulated according totechniques known in the art using suitable dispersing or wetting agentsand suspending agents. The sterile injectable preparation may also be asterile injectable solution or suspension in a non-toxic parenterallyacceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, such as carboxymethyl cellulose or similar dispersingagents that are commonly used in the formulation of pharmaceuticallyacceptable dosage forms including emulsions and suspensions. Othercommonly used surfactants, such as Tweens, Spans and other emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms may also be used for the purposes of formulation.

The pharmaceutically acceptable compositions of this invention may beorally administered in any orally acceptable dosage form including, butnot limited to, capsules, tablets, aqueous suspensions or solutions. Inthe case of tablets for oral use, carriers commonly used include lactoseand corn starch. Lubricating agents, such as magnesium stearate, arealso typically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, the pharmaceutically acceptable compositions of thisinvention may be administered in the form of suppositories for rectaladministration. These can be prepared by mixing the agent with asuitable non-irritating excipient that is solid at room temperature butliquid at rectal temperature and therefore will melt in the rectum torelease the drug. Such materials include cocoa butter, beeswax andpolyethylene glycols.

The pharmaceutically acceptable compositions of this invention may alsobe administered topically, especially when the target of treatmentincludes areas or organs readily accessible by topical application,including diseases of the eye, the skin, or the lower intestinal tract.Suitable topical formulations are readily prepared for each of theseareas or organs.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutically acceptable compositionsmay be formulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutically acceptable compositions canbe formulated in a suitable lotion or cream containing the activecomponents suspended or dissolved in one or more pharmaceuticallyacceptable carriers. Suitable carriers include, but are not limited to,mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax,cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutically acceptable compositions may beformulated, e.g., as micronized suspensions in isotonic, pH adjustedsterile saline or other aqueous solution, or, preferably, as solutionsin isotonic, pH adjusted sterile saline or other aqueous solution,either with or without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutically acceptablecompositions may be formulated in an ointment such as petrolatum. Thepharmaceutically acceptable compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

Most preferably, the pharmaceutically acceptable compositions of thisinvention are formulated for oral administration.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, dissolving orsuspending the compound in an oil vehicle accomplishes delayedabsorption of a parenterally administered compound form. Injectabledepot forms are made by forming microencapsule matrices of the compoundin biodegradable polymers such as polylactide-polyglycolide. Dependingupon the ratio of compound to polymer and the nature of the particularpolymer employed, the rate of compound release can be controlled.Examples of other biodegradable polymers include poly(orthoesters) andpoly(anhydrides). Depot injectable formulations are also prepared byentrapping the compound in liposomes or microemulsions that arecompatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, eardrops, and eye drops are also contemplated asbeing within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

The compounds of the invention are preferably formulated in dosage unitform for ease of administration and uniformity of dosage. The expression“dosage unit form” as used herein refers to a physically discrete unitof agent appropriate for the patient to be treated. It will beunderstood, however, that the total daily usage of the compounds andcompositions of the present invention will be decided by the attendingphysician within the scope of sound medical judgment. The specificeffective dose level for any particular patient or organism will dependupon a variety of factors including the disorder being treated and theseverity of the disorder; the activity of the specific compoundemployed; the specific composition employed; the age, body weight,general health, sex and diet of the patient; the time of administration,route of administration, and rate of excretion of the specific compoundemployed; the duration of the treatment; drugs used in combination orcoincidental with the specific compound employed, and like factors wellknown in the medical arts.

The amount of the compounds of the present invention that may becombined with the carrier materials to produce a composition in a singledosage form will vary depending upon the host treated, the particularmode of administration. Preferably, the compositions should beformulated so that a dosage of between 0.01-100 mg/kg body weight/day ofthe inhibitor can be administered to a patient receiving thesecompositions.

Depending upon the particular condition, or disease, to be treated orprevented, additional therapeutic agents, which are normallyadministered to treat or prevent that condition, may also be present inthe compositions of this invention. As used herein, additionaltherapeutic agents that are normally administered to treat or prevent aparticular disease, or condition, are known as “appropriate for thedisease, or condition, being treated.” Examples of additionaltherapeutic agents are provided infra.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered in a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

Uses of the Compounds and Compositions of the Invention

In one aspect of the invention, the invention features a method oftreating or lessening the severity of a PI3K-mediated condition ordisease in the brain or spinal cord of a patient. The term“PI3K-mediated disease”, as used herein means any disease or otherdeleterious condition in which a PI3K isoform is known to play a role.In one embodiment, the PI3K isoform is PI3Kγ.

Accordingly, in one embodiment, the invention features a method oftreating a PI3Kγ-mediated disease of the central nervous system. Suchconditions include, without limitation, inflammatory diseases andautoimmune-related diseases of the central nervous system. Accordingly,the invention provides a method of treating or lessening the severity ofa disease of condition selected from an autoimmune disease or aninflammatory disease of the central nervous system of a patient,comprising administering to said patient a compound or composition ofthe invention.

In a further embodiment, the compound of the invention is selective forthe inhibition of the PI3Kγ-isoform. In one embodiment, compounds of theinvention are selective for the inhibition of the PI3K gamma isoformover the PI3K alpha isoform in an in vitro assay by at least 20-fold. Inanother embodiment, the PI3Kγ-selective compounds of the inventioninhibit the gamma isoform over each of the alpha, beta, and deltaisoforms in an in vitro assay by at least 3-fold. In another embodiment,the PI3Kγ-selective compounds of the invention inhibit the gamma isoformover each of the alpha, beta, and delta isoforms in an in vitro assay byat least 5-fold. In yet another embodiment, the PI3Kγ-selectivecompounds of the invention inhibit the gamma isoform over each of thealpha, beta, and delta isoforms in an in vitro assay by at least10-fold.

In another embodiment, the invention provides a method of treating orlessening the severity of an inflammatory or autoimmune disease ordisorder of the central nervous system. In another embodiment, theinvention provides a method of treating or lessening the severity of asymptom of an inflammatory or autoimmune disease or disorder of thecentral nervous system. In a further embodiment, the invention providesa method of treating neuroinflammation. Such diseases or disordersinclude, without limitation, multiple sclerosis, transverse myelitis,progressive multifocal leukoencephalopathy, meningitis, encephalitis,myelitis, encephalomyelitis, intracranial or intraspinal abscess,phlebitis or thrombophlebitis of intracranial venous sinuses,Parkinson's Disease, Alzheimer's Disease, Huntington's Disease, Pick'sDisease, amyotrophic lateral sclerosis, HIV type-I dementia,frontotemporal lobe dementia, or traumatic brain or spinal cord injury.

PI3Kγ has also been reported to be important in diseases or disorderscharacterized by the activation of resident cells such as microglia. SeeJin et al., 2010, Biochemical and Biophysical Research Communications399:458-464. Microglia may potentiate damage to blood-brain barrier(BBB) integrity and endanger neuronal survival through the release ofreactive oxygen species or pro-inflammatory cytokines and otherneurotoxins. Inhibition of microglial activation may protect the brainafter ischemic stroke, as well as other neurovascular disorders such asmultiple sclerosis.

Compounds or compositions of the invention may be administered with oneor more additional therapeutic agents, wherein the additionaltherapeutic agent is appropriate for the disease being treated and theadditional therapeutic agent is administered together with a compound orcomposition of the invention as a single dosage form or separately fromthe compound or composition as part of a multiple dosage form. Theadditional therapeutic agent may be administered at the same time as acompound of the invention or at a different time. In the latter case,administration may be staggered by, for example, 6 hours, 12 hours, 1day, 2 days, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, or 2 months.

Non-limiting examples of chemotherapeutic agents or otheranti-proliferative agents that may be combined with the compounds ofthis invention include taxanes, aromatase inhibitors, anthracyclines,microtubule targeting drugs, topoisomerase poison drugs, targetedmonoclonal or polytonal antibodies, inhibitors of a molecular target orenzyme (e.g., a kinase inhibitor), or cytidine analogues. In oneembodiment, the additional chemotherapeutic agent is amsacrine,anastrozole, asparaginase, Avastin™ (bevacizumab) azathioprine,bicalutamide, bleomycin, camptothecin, carmustine, chlorambucil,cyclophosphamide, cytarabine (araC), daunonibicin, dactinomycin,doxorubicin (adriamycin), epirubicin, epothilone, etoposide, exemestane,fludarabine, 5-fluorouracil (5-FU), flutamide, Gemzar™ (gemcitabine),Gleevec™ (imatanib), Herceptin™ (trastuzumab), idarubicin, ifosfamide,an interferon, an interleukin, irinotecan, letrozole, leuprolide,lomustine, lovastatin, mechlorethamine, megestrol, melphalan,6-mercaptopurine, methotrexate (MTX), minosine, mitomycin, mitoxantrone,navelbine, nocodazole, platinum derivatives such as cisplatin,carboplatin and oxaliplatin, raloxifene, tamoxifen, Taxotere™(docetaxel), Taxol™ (paclitaxel), teniposide, topotecan, tumor necrosisfactor (TNF), vinblastin, vincristin, vindesine, vinorelbine, orZoladex™ (goserelin). Another chemotherapeutic agent can also be acytokine such as G-CSF (granulocyte colony stimulating factor). In yetanother embodiment, a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, analog orderivative thereof, may be administered in combination with surgery,radiation therapy, or with standard chemotherapy combinations such as,but not restricted to, CMF (cyclophosphamide, methotrexate and5-fluorouracil), CAF (cyclophosphamide, adriamycin and 5-fluorouracil),AC (adriamycin and cyclophosphamide), FEC (5-fluorouracil, epirubicin,and cyclophosphamide), ACT or ATC (adriamycin, cyclophosphamide, andpaclitaxel), or CMFP (cyclophosphamide, methotrexate, 5-fluorouracil andprednisone).

Additional therapeutic agents also include those useful for treatingmultiple sclerosis (MS), such as, for example, beta interferon (e.g.,Avonex® and Rebif®), glatiramir (Copaxone®), Tysabri® (natalizumab),Betaseron® (IFN-beta), and mitoxantrone.

The invention provides a method of inhibiting PI3K kinase activity in abiological sample that includes contacting the biological sample with acompound or composition of the invention. The term “biological sample,”as used herein, means a sample outside a living organism and includes,without limitation, cell cultures or extracts thereof; biopsied materialobtained from a mammal or extracts thereof; and blood, saliva, urine,feces, semen, tears, or other body fluids or extracts thereof.Inhibition of kinase activity, particularly PI3K kinase activity, in abiological sample is useful for a variety of purposes known to one ofskill in the art. Examples of such purposes include, but are not limitedto, biological specimen storage and biological assays. In oneembodiment, the method of inhibiting PI3K kinase activity in abiological sample is limited to non-therapeutic methods.

Preparation of Compounds of the Invention

As used herein, all abbreviations, symbols and conventions areconsistent with those used in the contemporary scientific literature.See, e.g., Janet S. Dodd, ed., The ACS Style Guide: A Manual for Authorsand Editors, 2nd Ed., Washington, D.C.: American Chemical Society, 1997.The following definitions describe terms and abbreviations used herein:

ATP adenosine triphosphateBrine a saturated NaCl solution in waterCp*RuCl(cod)chloro(pentamethylcyclopentadienyl)(cyclooctadiene)ruthenium(II)DCM dichloromethaneDIEA diisopropylethylamineDMA dimethylacetamideDMAP 4-dimethylaminopyridineDMF dimethylformamideDMSO methylsulfoxidedppfPdCl₂ 1,1′-bis(diphenylphosphino)-ferrocenedichloro-palladium.dichloromethaneDTT dithiothreitolEDCI 1-ethyl-3-(3-dimethylaminopropyl)carbodiimideESMS electrospray mass spectrometryEt₂O ethyl etherEtOAc ethyl acetateEtOH ethyl alcoholHATU 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphateHEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acidHPLC high performance liquid chromatographyLC-MS liquid chromatography-mass spectrometrym-CPBA meta-chloroperbenzoic acidMe methylMeOH methanolMTBE methyl t-butyl etherMC methyl cellulose

NMP N-methylpyrrolidine

PBS phosphate buffered salinePd(Cy)₃Cl₂ dichloro-bis(tricyclohexylphosphoranyl)palladium(II)Ph phenylRT or rt room temperature (between 20° C. and 25° C.)tBu tertiary butylTCA trichloroacetic acidTHF tetrahydrofuranTEA triethylamine

General Synthetic Procedures

In general, the compounds of this invention may be prepared by methodsdescribed herein or by other methods known to those skilled in the art.

Example 1 General Preparation of the Compounds of Formula I

Compounds of formula I can be prepared as shown in Scheme 1, Routes A toK, wherein each of A, B, C, D, E, X¹, X², R¹, and R² is as describedelsewhere in the description and each of R³ and R⁴ is H. As shown inRoute A of the scheme, for compounds of formula I where X is CH, thearyl halide of formula A1 is treated with N-bromosuccinimide (NBS) toproduce bromoalkyl compounds of formula of formula A2. Subsequentreaction of the bromoalkyl compounds with amines of formula A3 produce5-haloisoindolinones of formula A4. Reaction of a compound of formula A4with a boronic acid or boronate of formula A5 in the presence of anappropriate palladium catalyst produces compounds of formula I, whereineach of X¹ and X² is CH. Procedures for preparing a boronate or boronicacid from aryl or heteroaryl halides are described in Boronic Acids,ISBN: 3-527-30991-8, Wiley-VCH, 2005 (Dennis G. Hall, editor). In oneexample, the halogen is bromine and the boronate is prepared by reactingthe aryl or heteroaryl bromide with4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane.In subsequent coupling reactions, boronates or boronic acids so formedcan be reacted with aryl or heteroaryl halides in the presence of1,1′-bis(diphenylphosphino)-ferrocene dichloro-palladium.dichloromethane(dppfPdCl₂).

As shown in Route B, the sequence of steps outlined above can be changedsuch that compounds of formula A1 are first reacted with compounds ofA5, followed by formation of alkyl bromides A7 and then reacting thealkyl bromides with amines of formula A3 to produce compounds of formulaI, wherein X is CH and R⁵ is hydrogen.

As shown in Route C, in order to prepare compounds of formula I, whereinX¹ is N, compounds for formula A8 can be reacted with1,3,5-trichloro-1,3,5-triazinane-2,4,6-trione to form chloroalkylcompounds of formula A9. Compounds of formula A9 are then treated withoxidizing agents such as meta-chloroperbenzoic acid to form the N-oxidesof formula A10, which are subsequently treated with phosphorusoxychloride to produce pyridyl chlorides of formula All. Reaction of acompound of formula All with an amine of formula A3 produces a5-chloroazaisoindolinone of formula A12. Reaction of a compound offormula A12 with a boronic acid or boronate of formula A5 in thepresence of an appropriate palladium catalyst produces a compound offormula I, wherein X¹ is N and X² is CH or C—CH₃.

Compounds of formula I, wherein X¹ is CH or N, X² is N and each of R³and R⁴ is hydrogen can be also be prepared in a manner similar to thatdescribed for Route C, as shown in Route D.

Compounds of formula I, wherein X¹ is CH, X² is N and each of R³ and R⁴is hydrogen can be prepared as shown in as shown in Route E.Accordingly, amines of formula R¹—NH₂ are reacted withhalomethylacetylene to produce compounds of formula A18, which can thenbe reacted with carboxyacetylenes to produce compounds of formula A19.Reaction with ethyl carbonisocyanatidate in the presence ofchloro(pentamethylcyclopentadienyl)(cyclooctadiene)ruthenium(II) producecompounds of formula A20. Acidic hydrolysis and decarboxylation of theethoxyester followed by formation of chloropyridine via phosphorousoxychloride produces compounds of formula A21. Reaction with boronatesof formula A4 under catalytic cross-coupling conditions producecompounds of formula I.

Compounds of formula I, wherein an aryl or heteroaryl halide is used tointroduce R¹, can be prepared as shown in as shown in Route F.Accordingly, amides of formula A23 (prepared by the aminolysis ofcompounds of formula A22) are reacted with aryl chlorides, arylbromides, aryl iodides, or aryl triflates in the presence of a copper orpalladium catalyst to produce compounds of formula I.

As shown in Route G, compounds of formula I, where R¹ is a substitutedpyrazole, can also be prepared by reacting compounds of formula A25under basic conditions with an alkylating agent of formula R^(1a)—X,where X is a halide or sulfonate leaving group.

As shown in Route H, reaction of pyrrolopyridine diones of formula A26with a reducing agent, such as Zn/acetic acid, produces alcohols offormula A27. These alcohols can be subsequently alkylated with analkylating agent (e.g., L is a leaving group such as sulfonate or halideand R^(3′) is an aliphatic, cycloaliphatic, or heterocyclic group) inthe presence of a base to product compounds of formula A28. Compounds offormula A28 are then treated with oxidizing agents such asmeta-chloroperbenzoic acid to form the N-oxides of formula A29, whichare subsequently treated with phosphorus oxychloride to produce pyridylchlorides of formula A30. Reaction with boronates or boronic acids offormula A5 in a Pd-mediated cross coupling reaction produce compounds offormula I, where R³ is an ether-linked moiety and R⁴ is hydrogen. When amixture of enantiomers or diastereomers is present, such mixtures can beseparated by resolution methods such as supercritical fluidchromatography.

As shown in Route I, reaction of pyrrolopyridine diones of formula A26with a Grignard reagent, followed by dehydration and hydrogenation ofthe intermediate compound having formula A31, produce compounds offormula A32. Compounds of formula A32 are then treated with oxidizingagents such as meta-chloroperbenzoic acid to form the N-oxides offormula A33, which are subsequently treated with phosphorus oxychlorideto produce pyridyl chlorides of formula A34. Reaction with boronates orboronic acids of formula A5 in a Pd-mediated cross coupling reactionproduce compounds of formula I as a mixture of enantiomers. Suchmixtures can be separated by resolution methods such as supercriticalfluid chromatography.

As shown in Route J, compounds of formula I where neither R³ or R⁴ ishydrogen can be prepared by treatment of compounds of formula A35 withstrong non-nucleophilic base followed by reaction with at least twoequivalents of alkylating agent.

As shown in Scheme 1, Route K, compounds of formula I where R³ and R⁴and the intervening carbon form a ring can be prepared by treatment ofcompounds of formula A35 with strong non-nucleophilic base followed byreaction with an equivalent of a bis-alkylating agent.

The following representative examples provide details for thepreparation of the compounds of the invention.

Example 2 Preparation of 3-ethoxy-2-methoxy-5-bromopyridine (Compound2004)

As shown in step 2-i of Scheme 2, to NaH (4.0 g, 60% in mineral oil, 0.1mol) in a 100 mL DMF suspension was added 10 mL of an absolute ethylalcohol (4.6 g, 0.1 mol)/DMF solution at RT. After the evolution ofhydrogen gas, the reaction mixture was stirred at RT for 30 minutes andthe resulting ethoxide solution transferred to a solution of3,5-dibromopyridine (11.84 g, 0.05 mol, obtained from Aldrich ChemicalCo.) in 100 mL DMF at 60° C. The reaction was stirred at 60° C. for 4hours and then allowed to come to RT. Brine and ethyl acetate were addedand the organics were partitioned, dried over MgSO₄, filtered, and thevolatiles removed under reduced pressure. The resulting crude materialwas purified by silica gel chromatography, with the desired producteluting with 20% ethyl acetate/hexanes to give 3-bromo-5-ethoxypyridine(Compound 2001, 4.25 g, 42% yield): ¹H NMR (CDCl₃) δ 8.3 (dd, 2H), 7.4(d, 1H), 4.12 (q, 2H), 1.45 (t, 3H). 3-Benzyloxy-5-bromopyridine wasprepared by an analogous procedure: ¹H NMR (CDCl₃) δ 8.33 (d, 2H),7.5-7.35 (m, 6H), 5.15 (s, 2H).

Alternatively, as shown in step 2-ii of Scheme2,3-bromo-5-hydroxypyridine (100 mg, 0.57 mmol, obtained from AldrichChemical Co.) was diluted with DMF (3 mL). Potassium carbonate (158.8mg, 1.15 mmol) was added, followed by the addition of bromoethane (62.6mg, 42.6 μL, 0.57 mmol). The mixture was warmed to 60° C. and stirredovernight. After cooling, the mixture was dissolved in ethyl acetate andwashed with 2 M NaOH, followed by water. The organics were dried oversodium sulfate, filtered, and the volatiles removed under reducedpressure. The resulting crude 3-bromo-5-ethoxypyridine (Compound 2001)was used without further purification. The following compounds were madeby analogous procedures: 3-bromo-5-propoxypyridine, ESMS (M+H)218.19/216.19; 3-bromo-5-butylpyridine, ESMS (M+H) 230.22/232.22;3-bromo-5-(cyclohexylmethoxy)pyridine, ESMS (M+H) 270.2/272.22;3-(2-fluoroexthoxy)-5-bromopyridine, ESMS (M+H) 220.14/222.14;3-(2,2-difluoroexthoxy)-5-bromopyridine; and3-(2-ethylbutoxy)-5-bromopyridine, ESMS (M+H) 258.33/256.33.

As shown in step 2-iii of Scheme 2,3-chloroperoxybenzoic acid (9.426 g,42.06 mmol) was added to 3-bromo-5-methoxypyridine (4.25 g, 21 mmol) in200 mL of DCM at RT. The reaction was stirred overnight and the mixturewas washed with 200 mL of 2 N NaOH and 2×200 mL brine. The organic phasewas dried over MgSO₄, filtered and the volatiles removed under reducedpressure to provide 3-bromo-5-ethoxypyridine, 1-oxide (Compound 2002,4.4 g): ¹H NMR (CDCl₃): δ 8.05 (s, 1H), 7.9 (s, 1H), 7.0 (s, 1H), 4.12(q, 2H), 1.45 (t, 3H).

As shown in step 2-iv of Scheme 2, phosphorous oxychloride (48.02 g,403.6 mmol) was added to 3-bromo-5-ethoxypyridine, 1-oxide (4.4 g, 20.18mmol) in 700 mL of DCM at RT. The reaction mixture was stirred at RTovernight. After the addition of brine, the organics were partitioned,dried over MgSO₄, filtered, and the filtrate concentrated under reducedpressure. The product was purified by filtering the concentrate througha pad of silica gel and eluting the pad with ethyl acetate. Thevolatiles were removed under reduced pressure to provide5-bromo-2-chloro-3-ethoxypyridine (Compound 2003, 4.3 g, 85.6%): ¹H NMR(CDCl₃) δ 8.1 (s, 1H), 7.32 (s, 1H), 4.15 (q, 2H), 1.6 (t, 3H).

As shown in step 2-v of Scheme 2, 40.51 mL of a 25% MeONa/MeOH solutionwas added to 5-bromo-2-chloro-3-ethoxypyridine (4.3 g, 17.27 mmol). Thereaction mixture was refluxed for 2 hours. After cooling, ethyl acetateand brine were added to the mixture. The organic phase was dried withMgSO₄, filtered, and evaporated under reduced pressure. Afterpurification via silica gel chromatography,5-bromo-3-ethoxy-2-methoxypyridine (Compound 2004, 2.1 g, 50% yield) wasobtained: ¹H NMR (CDCl₃) δ 7.8 (s, 1H), 7.15 (s, 1H), 4.1 (q, 2H), 4.0(s, 3H), 1.5 (t, 3H). The following compounds were synthesized by ananalogous procedure: 5-Bromo-3-isopropoxy-2-methoxypyridine: ¹H NMR(CDCl₃) δ 7.7 (s, 1H), 7.1 (s, 1H), 4.55-4.5 (m, 1H), 3.9 (s, 3H), 1.3(d, 6H); 5-bromo-2-ethoxy-3-methoxypyridine: ESMS (M+H) 232, 234;5-bromo-3-methoxy-2-propoxypyridine: ESMS (M+H) 246, 248;5-bromo-2-isopropoxy-3-methoxypyridine: ESMS (M+H) 246, 248;5-bromo-2-(2,2-difluoroethoxy)-3-methoxypyridine: ESMS (M+H) 268, 270;5-bromo-2,3-diethoxypyridine: ESMS (M+H) 246, 248;5-bromo-2-(2,2-difluoroethoxy)-3-ethoxypyridine: ESMS (M+H) 282, 284;5-bromo-3-ethoxy-2-propoxypyridine: ESMS (M+H) 260, 262;5-bromo-3-ethoxy-2-isopropoxypyridine: ESMS (M+H) 260, 262;5-bromo-3-(2-fluoroethoxy)-2-methoxypyridine: ESMS (M+H) 250, 252;5-bromo-2-methoxy-3-propoxypyridine: ESMS (M+H) 246, 248;5-bromo-2-methoxy-3-(2-methoxyethoxy)pyridine: ESMS (M+H) 262, 264;5-bromo-3-(2,2-difluoroethoxy)-2-methoxypyridine: ¹H NMR (CDCl₃) δ 7.9(d, 1H), 7.2 (d, 1H), 6.1 (tt, 1H), 4.4 (q, 2H), 4.2 (td, 2H), 1.4 (t,3H); 5-bromo-2-ethoxy-3-isopropoxypyridine: ¹H NMR (CDCl₃) δ 7.7 (d,1H), 7.1 (d, 1H), 4.4 (m, 1H), 4.3 (q, 2H), 1.3 (m, 9H);5-bromo-3-butoxy-2-methoxypyridine: ESMS (M+H) 260, 262;5-bromo-2-methoxy-3-(2,2,2-trifluoroethoxy)pyridine: ESMS (M+H) 286,288; and 5-bromo-2-ethoxy-3-(2,2,2-trifluoroethoxy)pyridine: ESMS (M+H)300, 302.

Also prepared by a procedure analogous to that shown in Scheme 2 were5-methoxy-3-bromopyridine, 5-difluoromethoxy-3-bromopyridine,2-amino-3-difluoromethoxy-5-bromopyridine,2,3-dimethoxy-5-bromopyridine, 2-ethoxy-3-methoxy-5-bromopyridine,2-(2-methoxyethoxy)-3-methoxy-5-bromopyridine,2-(2-ethoxyethoxy)-3-methoxy-5-bromopyridine,2-(2-propyloxyethoxy)-3-methoxy-5-bromopyridine,2-(2-ethoxybutoxy)-3-methoxy-5-bromopyridine,2-(2-(2-methoxyethoxy)ethoxy)-3-methoxy-5-bromopyridine,2,3-diethoxy-5-bromopyridine, 2-methoxy-3-propoxy-5-bromopyridine,2-methoxy-3-(2-methoxyethoxy)-5-bromopyridine,2-methoxy-3-(2-butoxyethoxy)-5-bromopyridine,2-methoxy-3-(3-methoxypropyloxy)-5-bromopyridine,2-methoxy-3-(tetrahydro-2H-pyran-4-yloxy)-5-bromopyridine, and2-methoxy-3-(tetrahydro-2H-pyran-4-yloxy)-5-bromopyridine.

Example 3(a) Preparation of5-bromo-3-(difluoromethoxy)-2-methoxypyridine (Compound 2010)

As shown in step 3(a)-i of Scheme 3(a), 2-chloro-3-hydroxypyridine(Compound 2005, 2.0 g, 15.4 mmol, obtained from Aldrich Chemical Co.)was dissolved in 40 mL of DMF and 5.0 mL of water along with sodiumchlorodifluoroacetate (4.71 g, 30.9 mmol, obtained from LancasterSynthesis, Inc.) and anhydrous potassium carbonate (2.56 g; 18.5 mmol).The reaction mixture was heated in an oil bath at 100° C. for 2 hours.Another equivalent of sodium chlorodifluoroacetate and 1.2 equiv. ofpotassium carbonate were added and heating continued for an additional2.0 hours. After this time, the reaction was cooled and the volatilesremoved under reduced pressure. The residue was partitioned betweenbrine and ethyl acetate and the organics washed once more with brine,dried over Na₂SO₄, filtered, and the volatiles removed under reducedpressure. The product was purified by silica gel chromatography, elutingwith a hexanes/DCM to DCM gradient, to produce2-chloro-3-(difluoromethoxy)pyridine as a white solid (Compound 2006,2.0 g, 72% yield): ESMS (M+H) 180; ¹H NMR (CDCl₃) δ 8.05 (m, 1H), 7.45(m, 1H), 6.90 (m, 1H), 6.60 (t, 1H; J=75 Hz), 4.01 (s, 3H).

As shown in step 3(a)-ii of Scheme 3(a), an excess of sodium metal wasdissolved into 20 mL anhydrous methanol and2-chloro-3-(difluoromethoxy)pyridine (2.0 g, 11.1 mmol) in anhydrousmethanol was added. The reaction mixture was stirred in a sealed vesselat 100° C. for 6 hours. The volatiles were removed under reducedpressure and the residue was partitioned between EtOAc and brine. Thebrine was extracted with EtOAc and the combined organics were dried overNa₂SO₄, filtered, and the volatiles removed under reduced pressure. Theproduct was purified by silica gel chromatography (DCM) to yield3-(difluoromethoxy)-2-methoxypyridine as a colorless oil (Compound 2007,1.1 g, 56% yield: ESMS (M+H) 176.

As shown in step 3(a)-iii of Scheme 3(a),3-(difluoromethoxy)-2-methoxypyridine (270 mg, 1.54 mmol) was dissolvedin 5 mL of DCM and BBr₃ (540 μL; 1275 mg; 4.10 mmol) in heptane wasadded. The reaction mixture was stirred for 10 minutes at RT under anatmosphere of nitrogen, brought to reflux, and then stirred anadditional 4 hours. The mixture was cooled and water was added to quenchthe reaction. The pH was adjusted to 7-8 with sodium bicarbonate, theorganics partitioned, and the aqueous layer saturated with NaCl andextracted twice more with DCM. The combined organics were dried overNa₂SO₄, filtered, and the volatiles removed under reduced pressure. Theproduct was purified by silica gel chromatography (DCM to 5% MeOH/DCMgradient) to yield 3-(difluoromethoxy)pyridin-2-ol as a white solid(Compound 2008, 986 mg, 97% yield): ESMS (M+H) 162.

As shown in step 3(a)-iv of Scheme 3(a), 3-(difluoromethoxy)pyridin-2-ol(986 mg; 6.12 mmol) was dissolved in 25 mL of glacial acetic acid andsodium acetate (79 mg; 9.6 mmol) was added. The mixture was cooled in anice bath and bromine (780 μL; 1.63 g; 10.22 mmol) in 10 mL of glacialacetic acid was added over 10 minutes. The reaction was stirred for 30minutes at 10-15° C. The volatiles were removed under reduced pressureand the residue was partitioned between brine/saturated sodium carbonatesolution and ethyl acetate. After the evolution of gas ceased, theorganic and aqueous layers were separated and the aqueous solutionextracted three additional times with EtOAc. The combined organics weredried over Na₂SO₄, filtered, and the volatiles removed under reducedpressure. The residue was purified twice by silica gel chromatography(first a DCM to 10% MeOH/DCM gradient then 1:1 EtOAc/hexanes) to provide5-bromo-3-(difluoromethoxy)pyridin-2-ol as a light yellow powder(Compound 2009, 810 mg, 55% yield): ESMS (M+H) 241.9/243.9; ¹H NMR(CDCl₃) δ 13.2 (br m, 1H), 7.44 (d, 1H, J=2.1 Hz), 7.18 (d, 1H, J=2.1Hz), 6.92 (t, 1H, J=75 Hz).

As shown in step 3(a)-v of Scheme 3(a),5-bromo-3-(difluoromethoxy)pyridin-2-ol (300 mg; 1.25 mmol) wasdissolved in 5 mL of chloroform. Silver carbonate (690 mg; 2.5 mmol) andmethyl iodide (780 μL; 1.77 g; 12.5 mmol) were added and the mixturestirred at RT overnight. The reaction mixture was filtered throughdiatomaceous earth, which was washed with additional CHCl₃. Thefiltrates were concentrated under reduced pressure to yield an oil whichwas purified by silica gel chromatography to yield5-bromo-3-(difluoromethoxy)-2-methoxypyridine as a white solid (Compound2010, 250 mg, 78% yield): ESMS (M+H) 254/256; ¹H NMR (CDCl₃) δ 8.08 (d,1H, J=2.1 Hz), 7.56 (d, 1H, J=2.1 Hz), 6.60 (t, 1H, J=75 Hz), 3.98 (s,3H).

Example 3(b) Preparation of 5-bromo-2-ethoxy-3-methoxypyridine (Compound2011) and 5-bromo-2,3-dimethoxypyridine (Compound 2012)

As shown in step 3(b)-i of Scheme 3(b),5-bromo-2-chloro-3-methoxypyridine (1.0 g, 4.5 mmol, prepared in thesame manner as Compound 2003 in Example 2 starting with3-bromo-5-methoxypyridine) was treated with a sodium ethoxide/ethanolsolution (5.05 mL, 21% w/v, 13.5 mmol) and the reaction mixturemicrowave irradiated at 100° C. for 20 minutes. Water was added and theethanol evaporated under reduced pressure. The resulting aqueoussolution was extracted with DCM and ether, followed by drying thecombined extracts over MgSO₄. After filtration, removal of the volatilesunder reduced pressure provided 5-bromo-2-ethoxy-3-methoxypyridine(Compound 2011), 0.72 g, 69% yield): ESMS (M+H) 232.32/234.23. As shownin step 3(b)-ii of Scheme 3(b), Compound 2012 (ESMS (M+H) 218.32/220.23)was prepared in the same manner as Compound 2011, using sodium methoxidein methanol instead of sodium ethoxide in ethanol.

Example 4(a) Preparation of3-ethoxy-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2014)

As shown in step 4(a)-i of Scheme 4(a), iodoethane (13.69 g, 7.021 mL,87.75 mmol) was added to 5-bromo-2-methylpyridin-3-ol (5.5 g, 29.25mmol) and K₂CO₃ (12.13 g, 87.75 mmol) in 200 mL DMF. The mixture wasstirred at 70° C. overnight and sat'd NaHCO3 was added to the mixture.The mixture was extracted with EtOAc (3×) and the combined organics werewashed with water (3×) and brine. After drying over sodium sulfate, themixture was filtered and concentrated under reduced pressure. Theresidue was purified by medium pressure silica gel chromatography (30%EtOAc in hexane) to yield 5-bromo-3-ethoxy-2-methylpyridine (Compound2013, 4.6 g, 65% yield): ESMS (M+H) 216.18; ¹H NMR (CDCl₃) δ 8.14 (d,J=1.9 Hz, 1H), 7.20 (d, J=1.8 Hz, 1H), 4.03 (q, J=7.0 Hz, 2H), 2.43 (s,3H), 1.47 (t, J=7.0 Hz, 3H)

As shown in step 4(a)-ii of Scheme 4(a),5-bromo-3-ethoxy-2-methylpyridine (4.16 g, 19.25 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(5.866 g, 23.10 mmol) and KOAc (5.668 g, 57.75 mmol) were mixed in 200ml, dioxane. The mixture was degased for 1 h with N₂, then1,1′-bis(diphenylphosphino)-ferrocene dichloro-palladium.dichloromethane(162.6 mg, 0.1925 mmol) was added and the mixture was heated at 80° C.under N₂ for 16 hours. After cooling to RT, MTBE was added to themixture, which was then was through Florisil®. The solvent was removedunder reduced pressure to obtain a grey solid, which was triturated withhexanes to afford3-ethoxy-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2014, 4.05 g, 80% yield): ¹H NMR (CDCl₃) δ 8.43 (d, J=1.1 Hz,1H), 7.40 (s, 1H), 4.03 (d, J=7.0 Hz, 2H), 2.51 (s, 3H), 1.46 (t, J=7.0Hz, 3H), 1.37 (s, 12H).

Example 4(b) Preparation ofN-ethyl-2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)nicotinamide(Compound 2016)

As shown in step 4(b)-i of Scheme 4(b), HATU (8.194 g, 2.55 mmol) andDIPEA (5.570 g, 7.507 mL, 43.10 mmol) was added to a solution of5-bromo-2-methoxypyridine-3-carboxylic acid (5 g, 21.55 mmol) in DMF (50mL). The resulting solution was stirred for 10 minutes followed by theaddition of ethanamine hydrochloric acid (1.757 g, 2.196 mL, 21.55mmol). The resulting solution was stirred at room temperature for 5hours. To the reaction mixture was added water (100 mL) and ethylacetate (100 mL). The organic layer was separated and dried overanhydrous sodium sulfate, filtered, and concentrated under reducedpressure. The crude residue was purified by silica gel chromatography(0-2% methanol in dichloromethane gradient) to produce5-bromo-N-ethyl-2-methoxynicotinamide as off white solid (Compound 2015,3.4 g): ¹H NMR (DMSO-d₆) δ 8.41 (d, J=2.5 Hz, 1H), 8.31 (s, 1H),8.20-8.13 (m, 1H), 3.95 (s, 3H), 3.35-3.23 (m, 2H), 1.11 (t, J=7.2 Hz,3H).

As shown in step 4(b)-ii of Scheme 4(b), a sealed tube was charged withBis(dipinacolato)diboron (3.332 g, 13.12 mmol),dichloro-bis(tricyclohexylphosphoranyl)palladium (484.2 mg, 0.6560mmol), KOAc (3.863 g, 39.36 mmol), and 2-methyltetrahydrofuran (52.46mL). The mixture was degassed for 10 min then heated in an oil bath for12 h at 125° C. The reaction was deemed complete by HPLC. Workup byfiltration through florisil and concentration in vacuo. Triturated theyellow oil with hexanes causing ppt'n of off white solid. Collectedunder vacuum filtration and dried under high vacuum to constant mass(4.7 g). ¹H NMR (300 MHz, DMSO) d 8.43 (d, J=2.0 Hz, 1H), 8.27 (d, J=2.0Hz, 1H), 8.23 (d, J=5.5 Hz, 1H), 3.98 (s, 3H), 3.35-3.25 (m, 2H), 1.26(s, 12H), 1.12 (t, J=7.2 Hz, 3H)

Example 5 Preparation of2-[4-[5-(5,6-dimethoxy-3-pyridyl)-1-oxo-isoindolin-2-yl]pyrazol-1-yl]acetonitrile(Compound 3)

As shown in step 5-i of Scheme 5, methyl 4-bromo-2-(bromomethyl)benzoate(Compound 2018, 2.08 g, 6.75 mmol; prepared by reacting1-(4-bromo-2-methylphenyl)ethanone with NBS), 1H-pyrazol-4-amine (561mg, 6.75 mmol), and DIEA (873 mg, 1.18 mL, 6.75 mmol) were combined inDMF (7.78 mL) and heated at 110° C. for 90 min. The reaction mixture wasdiluted with MeOH (60 mL) and the resulting white crystalline solid wascollected by filtration and dried under vacuum to give5-bromo-2-(1H-pyrazol-4-yl)isoindolin-1-one (Compound 2019, 1.21 g, 4.35mmol, 64% yield): ESMS (M+H) 279.99.

As shown in step 5-ii of Scheme5,5-bromo-2-(1H-pyrazol-4-yl)isoindolin-1-one (1.2 g, 4.32 mmol) wascombined with cesium carbonate (1.69 g, 5.18 mmol) in DMF (10 mL) in asealable tube and nitrogen gas was bubbled through the solution for 5minutes. 2-Iodoacetonitrile (1.08 g, 468 μL, 6.47 mmol) was added andthe tube was sealed and heated to 110° C. in an oil bath for 18 hours.Additional iodoacetonitrile added (0.5 mL) and the reaction mixture washeated for an additional 24 hours. The reaction mixture was poured intoH₂O/EtOAc and the resulting dark brown solid was collected byfiltration. The solid was washed with MeOH and then diethyl ether toprovide 2-[4-(5-bromo-1-oxo-isoindolin-2-yl)pyrazol-1-yl]acetonitrile(Compound 2020, 920 mg, 2.9 mmol, 67% yield): ESMS (M+H) 319.04; ¹H NMR(DMSO-d₆) δ 8.35 (s, 1H), 7.92 (m, 2H), 7.70 (m, 2H), 5.53 (s, 2H), 4.88(s, 2H).

As shown in step 5-iii of Scheme 5,2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine[Compound 2021, 376 mg, 1.42 mmol; prepared by reacting5-bromo-2,3-dimethoxypyridine with4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane],2-[4-(5-bromo-1-oxo-isoindolin-2-yl)pyrazol-1-yl]acetonitrile (450 mg,1.42 mmol), and cesium carbonate (925 mg, 2.84 mmol) were taken up inDMSO (7.5 mL) in a sealable tube. Nitrogen gas was bubbled through thesolution for 5 minutes, dppfPdCl₂ (141 mg, 0.17 mmol) added, and thevessel sealed. The reaction mixture was heated to 100° C. for 50 min.After cooling, the mixture was poured into EtOAc/H₂O, the resulting darksolid material filtered off, and the organics passed through a plug offlorisil. The filtrate was concentrated to a solid under reducedpressure, the residue was suspended in MeOH, and the solid collected byfiltration to give2-[4-[5-(5,6-dimethoxy-3-pyridyl)-1-oxo-isoindolin-2-yl]pyrazol-1-yl]acetonitrileas a solid (Compound 3, 323 mg, 57% yield): ESMS (M+H) 376.28; ¹H NMR(DMSO-d₆) δ 8.37 (s, 1H), 8.11 (d, J=1.8 Hz, 1H), 7.99 (s, 1H), 7.95 (s,1H), 7.84 (m, 2H), 7.64 (d, J=1.7 Hz, 1H), 5.55 (s, 2H), 4.93 (s, 2H),3.93 (s, 3H), 3.91 (s, 3H).

Example 6 Preparation of2-(4-(5-(5,6-dimethoxypyridin-3-yl)-7-methyl-1-oxoisoindolin-2-yl)-1H-pyrazol-1-yl)acetonitrile(Compound 48)

As shown in step 6-i of Scheme 6,2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2021, 920 mg, 3.47 mmol), methyl 4-bromo-2,6-dimethyl-benzoate(844 mg, 3.47 mmol), and Cs₂CO₃ (2.26 g, 6.94 mmol) were taken up inDMSO (12 mL) in a sealable tube. Nitrogen gas was bubbled through thesolution for 5 minutes, dppfPdCl₂ (141 mg, 0.174 mmol) added, and thevessel sealed. The reaction mixture was heated at 90° C. for 60 minutesunder an atmosphere of nitrogen. After cooling, the mixture was pouredinto EtOAc/water. The organics were washed with water, brine, passedthrough a plug of Florisil®, and concentrated under reduced pressure togive a solid. The solid was suspended in MeOH and collected byfiltration to provide methyl4-(5,6-dimethoxy-3-pyridyl)-2,6-dimethyl-benzoate (Compound 2022, 310mg). The filtrate was concentrated and purified by silica gelchromatography (0 to 50% EtOAc/hex) to provide an additional 400 mg ofCompound 2022 (total yield 710 mg, 2.4 mmol, 68% yield). This compoundwas used in subsequent reactions as is.

As shown in step 6-ii of Scheme 6, Compound 2022 (710 mg, 2.36 mmol) wasdissolved in CCl₄ (20 mL) and K₂CO₃ (651 mg, 4.71 mmol), NBS (461 mg,2.59 mmol), and benzoyl peroxide (57 mg, 0.24 mmol) were added. Thereaction mixture was heated to reflux for 4 hours. After cooling to roomtemperature, the reaction mixture was filtered and the solid washed withCCl₄. The filtrate was concentrated to an oil under reduced pressure andpurified by silica gel chromatography to provide methyl2-(bromomethyl)-4-(5,6-dimethoxy-3-pyridyl)-6-methyl-benzoate (Compound2023, 685 mg, about 70% pure). This compound was used in subsequentreactions as is.

As shown in step 6-iii of Scheme 6, 1H-pyrazol-4-amine (149 mg, 1.79mmol), methyl2-(bromomethyl)-4-(5,6-dimethoxy-3-pyridyl)-6-methyl-benzoate (680 mg,1.79 mmol) and DIEA (231 mg, 312 μL, 1.79 mmol) were combined in DMF (5mL), heated to 90° C. for 6 hours, and allowed to cool to roomtemperature over 16 hours. The reaction mixture was taken up inEtOAc/water and the organic layer washed with water, brine, dried, andconcentrated under reduced pressure to yield a foam. The foam wasrecrystallized in DCM/MeOH. The resulting solid was collected byfiltration, washed with DCM, and dried under vacuum to provide5-(5,6-dimethoxy-3-pyridyl)-7-methyl-2-(1H-pyrazol-4-yl)isoindolin-1-one(Compound 2024, 115 mg). The filtrate from the recrystallization wasconcentrated under reduced pressure and the residue purified by silicagel chromatography (20 to 100% EtOAc/hex) to yield an additional 88 mgof Compound 2024 (total yield 203 mg, 0.66 mmol, 32% yield): ESMS (M+H)351.26; ¹H NMR (DMSO-d₆) δ 12.83 (s, 1H), 8.10 (m, 2H), 7.88 (s, 1H),7.74 (s, 1H), 7.61 (s, 2H), 4.83 (s, 2H), 3.92 (s, 3H), 3.91 (s, 3H),2.72 (s, 3H).

As shown in step 6-iv of Scheme6,5-(5,6-dimethoxy-3-pyridyl)-7-methyl-2-(1H-pyrazol-4-yl)isoindolin-1-one(88 mg, 0.25 mmol) was dissolved in DMF (2 mL) with Cs₂CO₃ (123 mg, 0.38mmol). 2-Bromoacetonitrile (45 mg, 0.38 mmol) was added and the reactionmixture stirred overnight at room temperature. The reaction mixture wasdiluted with MeOH/H₂O/EtOAc and the resulting white solid was collectedby filtration; washed sequentially with water, MeOH, and Et₂O; and driedunder vacuum to provide2-(4-(5-(5,6-dimethoxypyridin-3-yl)-7-methyl-1-oxoisoindolin-2-yl)-1H-pyrazol-1-yl)acetonitrile(Compound 48, 30 mg, 0.077 mmol, 29% yield): ESMS (M+H) 390.29; ¹H NMR(DMSO-d₆) δ 8.34 (s, 1H), 8.10 (d, J=1.8 Hz, 1H), 7.95 (s, 1H), 7.76 (s,1H), 7.68-7.53 (m, 2H), 5.53 (s, 2H), 4.85 (s, 2H), 3.92 (s, 3H), 3.91(s, 3H), 2.72 (s, 3H).

Example 7 Preparation of2-(4-(7-chloro-5-(5,6-dimethoxypyridin-3-yl)-1-oxoisoindolin-2-yl)-1H-pyrazol-1-yl)acetonitrile(Compound 58)

As shown in step 7-i of Scheme 7,2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(503 mg, 1.9 mmol), methyl 4-bromo-2-chloro-6-methyl-benzoate (500 mg,1.9 mmol), and Cs₂CO₃ (1.24 g, 3.8 mmol) were combined in DMSO (7 mL) ina sealable tube. Nitrogen gas was bubbled through the solution for 5minutes, dppfPdCl₂ (78 mg, 0.1 mmol) added, and the vessel sealed. Thereaction mixture was heated at 90° C. for 60 minutes under an atmosphereof nitrogen. After cooling, the mixture was poured into EtOAc/water. Theorganic layer washed with water, brine, passed through a plug ofFlorisil®, and concentrated under reduced pressure. The resulting oilwhich was purified by silica gel chromatography (0 to 50% EtOAc/hexanes)to provide methyl 2-chloro-4-(5,6-dimethoxy-3-pyridyl)-6-methyl-benzoate(Compound 2025, 367 mg, 1.14 mmol, 60% yield): ESMS (M+H) 322.12.

As shown in step 7-ii of Scheme 7, methyl2-chloro-4-(5,6-dimethoxy-3-pyridyl)-6-methyl-benzoate (365 mg, 1.13mmol) was dissolved in CCl₄ (20 mL) and NBS (162 mg, 0.907 mmol) added.The reaction mixture heated to reflux for 3 hours, at which time K₂CO₃(150 mg) and additional NBS (60 mg) were added. The reaction mixture washeated an additional 6 hours. At this time, HPLC analysis indicated thatabout 30% of the starting material (Compound 2025) had been converted toCompound 2026. Additional NBS (60 mg) was added and the reaction mixturerefluxed for an additional 6 hours, followed by allowing the mixture tostand at room temperature overnight. At this time, HPLC analysisindicated that about 50% of the starting material had been converted toproduct. The reaction mixture was filtered, washed with CCl₄, and thefiltrate concentrated under reduced pressure to give crude2-(4-(7-chloro-5-(5,6-dimethoxypyridin-3-yl)-1-oxoisoindolin-2-yl)-1H-pyrazol-1-yl)acetonitrile(Compound 2026) as an oil, which was used in subsequent reactions as is.

As shown in step 7-iii of Scheme 7, Compound 2026 obtained from step7-ii was dissolved in DMF (5 mL) and 2-(4-aminopyrazol-1-yl)acetonitrile(100 mg, 0.82 mmol) and DIEA (226 mg, 304 μL, 1.75 mmol) were added. Thereaction mixture was heated to 80° C. for 4 hours, cooled to roomtemperature, and allowed to stand overnight. The mixture was poured intoEtOAc/water (1:1 160 mL). The organic layer was washed with water,dried, and concentrated under reduced pressure to a yield a residuewhich solidified when treated with MeOH (20 mL). The solid was collectedby filtration, taken up in EtOAc (30 mL), and heated to reflux. Aftercooling to room temperature, the crystalline product was collected byfiltration dried in vacuo to provide2-(4-(7-chloro-5-(5,6-dimethoxypyridin-3-yl)-1-oxoisoindolin-2-yl)-1H-pyrazol-1-yl)acetonitrile(Compound 58, 68 mg, 0.17 mmol, 15% yield): ESMS (M+H) 410.3; ¹H NMR(DMSO-d₆) δ 8.36 (s, 1H), 8.16 (d, J=1.8 Hz, 1H), 7.97 (m, 2H), 7.92 (s,1H), 7.69 (s, 1H), 5.56 (s, 2H), 4.90 (s, 2H), 3.93 (s, 3H), 3.92 (s,3H).

Example 8 Preparation of2-(4-(2-(5,6-dimethoxypyridin-3-yl)-5-oxo-5H-pyrrolo[3,4-b]pyridin-6(7H)-yl)-1H-pyrazol-1-yl)acetonitrile(Compound 25)

As shown in step 8-i of Scheme 8, according to the procedure ofInternational Patent Application Publication No. WO2006/095159, amixture of ethyl 2-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (5.92g, 32.6 mmol) in phosphorous oxychloride (45 mL) was heated at 90° C.for 1 hour. After cooling, the reaction mixture was concentrated underreduced pressure and ice water was added to the residue, followed byaddition 28% ammonium hydroxide to adjust the pH to 7. The resultingwhite solid was collected by filtration, washed with ice water, anddried under high vacuum to give ethyl 6-chloro-2-methylnicotinate(Compound 2027, 6.18 g, 94.7% yield): ESMS (M+1) 200.19; ¹H NMR (CDCl₃)δ 8.18 (d, J=8.2 Hz, 1H), 7.27 (d, J=8.2 Hz, 1H), 4.40 (q, J=7.1 Hz,2H), 2.84 (s, 3H), 1.42 (t, J=7.4, 3H).

As shown in step 8-ii of Scheme 8, a mixture of ethyl6-chloro-2-methylpyridine-3-carboxylate (Compound 2027, 4.0 g, 20 mmol),2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2021, 5.84 g, 22 mmol), Pd(PPh₃)₄ (1.15 g, 1 mmol), and sodiumcarbonate (6.37 g, 60 mmol) in a mixture of acetonitrile/water (3:1, 90mL) was heated at 90° C. under an atmosphere of nitrogen for 4 hours.After cooling, the volatiles were removed under reduced pressure and theresidue dissolved in DCM. After washing with water, the organic phasewas dried (Na₂SO₄), filtered, and concentrated under reduced pressure.The residue was purified by silica gel chromatography (0-20%EtOAc/hexanes) to give ethyl5′,6′-dimethoxy-6-methyl-2,3′-bipyridine-5-carboxylate (Compound 2028,5.8 g, 95.7% yield): ESMS (M+1) 303.41; ¹H NMR (CDCl₃) δ 8.27 (d, J=1.8Hz, 1H), 8.17 (d, J=8.2 Hz, 1H), 7.83 (d, J=1.8 Hz, 1H), 7.52 (d, J=8.3Hz, 1H), 4.32 (q, J=7.1 Hz, 2H), 4.01 (s, 2H), 3.93 (s, 3H), 2.83 (s,3H), 1.34 (t, J=7.1 Hz, 3H).

As shown in step 8-iii of Scheme 8, to a solution of Compound 2029 (4.4g, 14.6 mmol) in CCl₄ (75 mL) was added 2,2′-azobis(isobutyronitrile)(AIBN, 239 mg, 1.46 mmol) and NBS (1.7 g, 9.55 mmol). The mixture wasstirred at 80° C. for 1.5 hours under an atmosphere of nitrogen. Thereaction mixture was filtered and concentrated. The residue was purifiedby silica gel chromatography (20% EtOAc/hexanes) to give a mixture ofstarting material and ethyl5′,6′-dimethoxy-6-bromomethyl-2,3′-bipyridine-5-carboxylate (Compound2029, 4.44 g, about 60% pure): ESMS (M+1) 381.4, 383.19. The product wasused as is in subsequent reactions.

As shown in step 8-iv of Scheme 8, a solution of the above mixture(Compound 2029, 2.2 g, about 60% pure) in DMF (40 mL) at 0° C. was addeddropwise over 2 hours to a suspension of2-(4-amino-1H-pyrazol-1-yl)acetonitrile (844 mg, 6.9 mmol) and sodiumcarbonate (732 mg, 6.9 mmol) in DMF (20 mL). After addition wascomplete, the reaction mixture was stirred at 0° C. for 2 hours andheated at 80° C. for 15 hours. Additional sodium carbonate (732 mg) wasadded and the reaction mixture was heated at 90° C. for an additional 7hours. After cooling, the mixture was poured into water and aprecipitate formed. The solid was collected by filtration, washed withmethyl t-butyl ether, and dried under high vacuum to provide2-(4-(2-(5,6-dimethoxypyridin-3-yl)-5-oxo-5H-pyrrolo[3,4-b]pyridin-6(7H)-yl)-1H-pyrazol-1-yl)acetonitrile(Compound 25, 450 mg).

Example 9 Preparation of2-(5,6-dimethoxypyridin-3-yl)-4-methoxy-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 88)

As shown in step 9-i of Scheme 9, ethyl4,6-dihydroxy-2-methylpyridine-3-carboxylate (Accla Biochem Inc.) wassuspended in 50 mL of POCl₃ and heated to 90° C. under a nitrogenatmosphere for 5 hours. The reaction mixture was cooled and concentratedunder reduced pressure. Ice was added to the dark oil with stirringfollowed by the addition of ethyl acetate and water. The organics werewashed with water, brine, and dried over sodium sulfate. Afterfiltration, the volatiles were removed under reduced pressure and thecrude product purified by silica gel chromatography to yield ethyl4,6-dichloro-2-methylnicotinate as a pale yellow oil (Compound 2030, 62%yield): ESMS (M+H) 234/236/238; ¹H NMR (CDCl₃) δ 7.27 (s, 1H), 4.4(quart, 2H), 2.55 s, 3H), 1.41 (t, 3H).

As shown in step 9-ii of Scheme 9, Compound 2030 (500 mg, 2.14 mmol) andof 2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2021, 566 mg, 2.14 mmol) were dissolved into 10 mL of DME andflushed with nitrogen for 5 minutes. Tetrakistriphenylphosphinepalladium(0) (250 mg, 0.2136 mmol) was added while continuing the flowof nitrogen. A solution of saturated aqueous 2M K₂CO₃ (2.2 mL) (flushedwith nitrogen) was added and the mixture was heated to 70° C. for 1.5hours. The volatiles were removed under reduced pressure and the residuetreated with water and 2 mL of 1.0N HCl. A precipitate formed which waspartitioned between EtOAc and water. The organics were washed withwater, brine, dried over sodium sulfate, and the volatiles removed underreduced pressure. The crude product was purified by silica gelchromatography (DCM-25% EtOAc/DCM) to provide ethyl4-chloro-5′,6′-dimethoxy-6-methyl-2,3′-bipyridine-5-carboxylate(Compound 2031, 650 mg, 90% yield) of a white-beige solid: ESMS (M+1)337/339; ¹H NMR (CDCl₃) δ 8.26 (d, 1H J=2 Hz), 7.77 (d, 1H, J=2 Hz),7.56 (s, 1H), 4.48 (quart, 2H), 4.08 (s, 3H), 4.00 (s, 3H), 2.63 (s,3H), 1.43 (t, 3H).

As shown in step 9-iii of Scheme 9, Compound 2031 (650 mg, 1.93 mmol)was dissolved in anhydrous methanol and to it was added 3.0 mL offreshly made 2.54M sodium methoxide. The reaction was heated under anitrogen atmosphere at 60° C. for 16 hours. After cooling to roomtemperature, the volatiles were removed under reduced pressure and theresidue was partitioned between water and EtOAc. The organics werewashed with water, brine, dried over sodium sulfate, filtered, and thevolatiles removed under reduced pressure. Purification by silica gelchromatography provided ethyl4,5′,6′-trimethoxy-6-methyl-2,3′-bipyridine-5-carboxylate (Compound2032, 200 mg, 32% yield) as a of white solid: ESMS (M+1) 319; ¹H NMR(CDCl₃) δ 8.23 (d, 1H, J=2 Hz), 7.8 (d, 1H, J=2 Hz), 7.06 (s, 1H), 4.07(s, 3H), 3.99 (s, 3H), 3.94 (s, 3H).

As shown in step 9-iv of Scheme 9, Compound 2032 (200 mg, 0.628 mmol)and NBS (112 mg, 0.63 mmol) were added to 15 mL of CCl₄ and the solutionpurged with nitrogen for 5 minutes. Benzoyl peroxide (20 mole %) wasadded and the reaction mixture heated at 65° C. under nitrogen for 16hours. An additional equivalent of NBS and 0.3 equivalents of benzoylperoxide were added and heating continued for an additional hour.Potassium carbonate (1.0 g) was added as an acid scavenger and heatingcontinued for an additional 24 hours. The reaction mixture was cooledand filtered through a cotton plug and the volatiles removed underreduced pressure. The residue was taken up in DCM and passed through aplug of silica gel, which was eluted with additional DCM. Elution withEtOAc recovered crude product, which was further purified by silica gelchromatography (DCM-1:1 EtOAc/DCM to give ethyl6-(bromomethyl)-4,5′,6′-trimethoxy-2,3′-bipyridine-5-carboxylate(Compound 2033, 87 mg of white solid: ESMS (M+1) 397/399.

As shown in step 9-v of Scheme 9, Compound 2033 (100 mg, 0.252 mmol),1-(2,2,2-trifluoroethyl)-4-aminopyrazole (42 mg, 0.25 mmol), and DIEA(65 mg, 0.5 mmol) were dissolved in 4 mL of DMF and heated for 3 hoursat 110° C. The reaction mixture was cooled, diluted with water, andextracted with EtOAc. The organics were washed with water, brine, driedover sodium sulfate, and solvent removed under reduced pressure. Thecrude product was passed through a plug of silica gel, which was elutedwith 5% EtOH/EtOAc. Purification of the filtrate by silica gelchromatography (5% MeOH/DCM-7.5% MeOH/DCM) gave2-(5,6-dimethoxypyridin-3-yl)-4-methoxy-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 88, 11 mg, 9% yield) as a beige solid: ESMS (M+1) 450.

Example 10 Preparation of4-chloro-2-(5,6-dimethoxypyridin-3-yl)-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 82) and2-(5,6-dimethoxypyridin-3-yl)-4-ethyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 182)

As shown in steps 10-i and 10-ii of Scheme 10, Compound 2031 wasconverted to Compound 82 in a manner similar to that in the conversionof Compound 2032 to Compound 88.

As shown in step 10-iii Scheme 10,4-Chloro-2-(5,6-dimethoxypyridin-3-yl)-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 82, 75 mg, 0.165 mmol and a degassed solution of 2M aqueoussodium carbonate (826 uL, 1.65 mmol] were suspended in 10 mL of toluene.The reaction mixture was purged with nitrogen for 5 minutes andtetrakistriphenylphosphine palladium(0) (38 mg, 0.033 mmol] was added,followed by the addition of 2 M triethylborate solution in THF (992 μL,1.0 mmol). The reaction vessel was sealed and heated for 14 hours at 80°C. The reaction mixture was cooled and the volatiles removed underreduced pressure. The residue was dissolved in EtOAc/DCM 1:1 andfiltered. The filtrate was concentrated under reduced pressure. Theproduct was purified by preparative thin layer chromatography produced2-(5,6-dimethoxypyridin-3-yl)-4-ethyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 182, 1.1 mg): ESMS (M+1) 448.

Example 11 Preparation of2-(5,6-dimethoxypyridin-3-yl)-3-methyl-6-(1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 238)

As shown in step 11-i of Scheme 11, 2,5-dimethylnicotinic acid (Compound2034, 519 mg, 3.43 mmol) was dissolved in 1,1,1-triethoxyethane (5.57 g,6.29 mL, 34.3 mmol) in a microwave vial. The reaction mixture was heatedto 150° C. for 5 minutes. After dilution with 30 mL of DCM, the organicswere washed with 10 mL of saturated NaHCO₃. The organic layer was passedthrough a phase separator and then concentrated under reduced pressureto give ethyl 2,5-dimethylnicotinate (Compound 2035, 390 mg, 63% yield)as a yellow oil: ESMS (M+1) 179.89; ¹H NMR (300 MHz, DMSO-d₆) δ 8.46 (d,J=1.9 Hz, 1H), 7.98 (d, J=1.9 Hz, 1H), 4.31 (q, J=7.1 Hz, 2H), 2.65 (s,3H), 2.31 (s, 3H), 1.32 (t, J=7.1 Hz, 3H).

As shown in step 11-ii of Scheme 11, Compound 2035 (354 mg, 1.98 mmol)and 1,3,5-trichloro-1,3,5-triazinane-2,4,6-trione (689 mg, 2.96 mmol)were combined in DCM (1.8 mL). After stirring 18 hours at roomtemperature, the mixture was diluted with 20 mLs each of saturatedsodium carbonate and dichloromethane. The organics were separated,washed with saturated sodium carbonate, washed with brine, dried oversodium sulfate, filtered, and concentrated under reduced pressure togive ethyl 2-(chloromethyl)-5-methylnicotinate (Compound 2036, 465 mg)as a pale yellow oil: ESMS (M+1) 213.86.

As shown in step 11-iii of Scheme 11, Compound 2036 (465 mg, 2.18 mmol)was placed in a 40 ml vial, diluted with DCM (4.35 mL), and3-chlorobenzenecarboperoxoic acid (m-CPBA, 551 mg, 2.39 mmol) was addedat room temperature with stirring. After 18 hours, the mixture wasdiluted with 30 mL of DCM, washed with saturated sodium carbonate (3×5mL), and washed with brine. The organics were passed through a phaseseparator and concentrated to dryness under reduced pressure to giveethyl 2-(chloromethyl)-3-(ethoxycarbonyl)-5-methylpyridine 1-oxide(Compound 2037, 318 mg, 1.38 mmol, 63% yield): ESMS (M+1) 230.14. Thismaterial was used as is in subsequent reactions.

As shown in step 11-iv of Scheme 11, Compound 2037 (318 mg, 1.385 mmol)was dissolved in phosphorus oxychloride (4.25 g, 2.58 mL, 27.7 mmol).The reaction mixture was heated to 90° C. under an atmosphere ofnitrogen for 18 hours. The mixture was concentrated to dryness underreduced pressure, diluted with 5 mL of DCM, and washed with 5 mL ofwater. The organics were passed through a phase separator and thevolatiles removed under reduced pressure. Purification by silica gelchromatography gave ethyl 6-chloro-2-(chloromethyl)-5-methylnicotinate(Compound 2038, 78 mg, 0.314 mmol, 22.7%): ESMS (M+1) 248.17; ¹H NMR(300 MHz, DMSO-d₆) δ 8.29 (s, 1H), 5.00 (s, 2H), 4.51-4.23 (m, 2H), 2.38(s, 3H), 1.35 (t, J=7.1 Hz, 3H).

As shown in step 11-v of Scheme 11, Compound 2038 (76 mg, 0.306 mmol)was dissolved in 1 mL of DMF and added dropwise to a stirring solutionof 1H-pyrazol-4-amine (63.6 mg, 0.766 mmol) and DIEA (59.4 mg, 801 μL,0.46 mmol) in 1 mL of DMF. The reaction was stirred at room temperaturefor 2 hours and then heated overnight at 80° C. After the addition of 10mL of methanol, the mixture was allowed to cool to produce a solid. Thesolid was collected by filtration and washed with 3 mL of methanol. Thesolid was dried overnight under high vacuum to give2-chloro-3-methyl-6-(1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2039, 35 mg, 0.141 mmol, 46% yield). ESMS (M+1) 249.08.

As shown in step 11-vi of Scheme 11,2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2028, 45 mg, 0.169 mmol), 1 M sodium carbonate (281 μL, 0.281mmol), and Compound 2039 (35 mg, 0.141 mmol) were taken up in 3 mL DMFas a slurry. The mixture was degassed with nitrogen for 30 minutes andPd(PPh₃)₄ (32.5 mg, 0.028 mmol) was added. The mixture was degassed withnitrogen for another 5 minutes and then heated for 18 hours at 80° C. ina sealed vial. Additional methanol was added followed by dilution withDCM. The organics were washed with a solution of saturated Na₂CO₃,passed through a phase separator, and concentrated under reducedpressure to dryness. The product was purified by reversed-phase HPLC(10-90% acetonitrile/water) to give2-(5,6-dimethoxypyridin-3-yl)-3-methyl-6-(1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 238, 18.8 mg, 0.052 mmol, 37% yield); ESMS (M+1) 352.26; ¹HNMR (300 MHz, CDCl₃) δ 8.10 (d, J=11.8 Hz, 3H), 7.97 (d, J=1.9 Hz, 1H),7.34 (d, J=1.8 Hz, 1H), 4.83 (s, 2H), 4.10 (s, 3H), 3.96 (s, 3H), 2.54(s, 3H).

Example 12 Preparation of2-(5,6-dimethoxypyridin-3-yl)-4-methyl-6-(1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 135)

As shown in step 12-i of Scheme 12, to a degassed mixture of Compound2021 (111 mg, 0.42 mmol), sodium carbonate (97 mg, 0.91 mmol) and2-chloro-4-methyl-6-(1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2040, 104 mg, 0.42 mmol; prepared from ethyl6-chloro-2-(chloromethyl)-4-methylnicotinate in a manner similar to thatof the preparation of Compound 2039 in step 11-v of Example 11) inDMF/acetonitrile/water (1:1:0.5) was added Pd(PPh₃)₄ (50 mg, 0.04 mmol).The reaction mixture was heated in a sealed tube at 90° C. for 48 hours.Water (5 mL) was added and the mixture stirred at RT for 30 minutes.After filtration, the collected solid was washed with MeOH and EtOAc,sonicated in EtOAc, then collected by filtration to give2-(5,6-dimethoxypyridin-3-yl)-4-methyl-6-(1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 135, 100 mg, 66% yield) as a pale green solid: ESMS (M+H)352.4; ¹H NMR (300 MHz, DMSO-d₆) δ 12.88 (s, 1H), 8.54 (d, J=1.9 Hz,1H), 8.29-7.66 (m, 4H), 4.90 (s, 2H), 3.93 (d, J=10.5 Hz, 6H), 2.72 (s,3H).

Example 13 Preparation of2-(5,6-dimethoxypyridin-3-yl)-4-methyl-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2043)

As shown in step 13-i of Scheme 13, ethyl6-chloro-2-(chloromethyl)-4-methylnicotinate (Compound 2041, 5.11 g,20.6 mmol; prepared from 2,5-dimethylnicotinic acid in a manner similarto the preparation of Compound 2038 in Example 11) was dissolved inmethanol (30.6 mL). 7M ammonia/MeOH (21.3 mL, 149 mmol) was addedfollowed by the addition of ammonium hydroxide (18.7 g, 20.8 mL, 533mmol). The reaction mixture was stirred overnight at room temperatureand the precipitate that had formed was collected by filtration anddried under high vacuum to provide2-chloro-4-methyl-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one (Compound2042, 2.6 g, 14.2 mmol, 69% yield): ESMS (M+1) 183.29; ¹H NMR (300 MHz,CDCl₃) δ 8.74 (s, 1H), 7.44 (d, J=0.5 Hz, 1H), 4.35 (s, 2H), 2.60 (s,3H).

As shown in step 13-ii of Scheme 13,2-chloro-4-methyl-6,7-dihydropyrrolo[3,4-b]pyridin-5-one (2.54 g, 13.91mmol), 1M sodium carbonate (27.82 mmol), and2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(4.43 g, 16.7 mmol) were slurried in DMF (216 mL). The reaction mixturewas flushed with nitrogen for 30 minutes. Pd(PPh₃)₄ (1.607 g, 1.391mmol) was added and the nitrogen flush was continued for another 5minutes. The reaction mixture was then heated at 80° C. for 16 hours.The mixture was diluted with 1 L ethyl acetate and 350 mL saturatedNaHCO₃. A precipitate formed in the separatory funnel which wascollected by filtration and washed with EtOAc, water, and ethyl ether.Drying the solid overnight under high vacuum gave2-(5,6-dimethoxy-3-pyridyl)-4-methyl-6,7-dihydropyrrolo[3,4-b]pyridin-5-one(Compound 2043, 3.82 g, 13.38 mmol, 96% yield): ESMS (M+H) 286.29; ¹HNMR (300 MHz, DMSO-d₆) δ 8.63 (s, 1H), 8.49 (d, J=1.9 Hz, 1H), 8.06-7.82(m, 2H), 4.41 (s, 2H), 3.94 (s, 3H), 3.90 (s, 3H), 2.67 (s, 3H).

Example 14 Preparation of2-(5,6-dimethoxypyridin-3-yl)-6-(5-methylthiophen-2-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 252)

As shown in step 14-i of Scheme 14,2-(5,6-dimethoxy-3-pyridyl)-6,7-dihydropyrrolo[3,4-b]pyridin-5-one(Compound 2044, 100 mg, 0.37 mmol; prepared via the aminolysis ofCompound 2029 as shown in step 13-i of Example 13),2-iodo-5-methyl-thiophene (99 mg, 54 μL, 0.44 mmol), and cesiumcarbonate (240 mg, 0.737 mmol) were weighed into a small screw top tube.The reaction mixture was flushed with nitrogen for 15 minutes. CuI (14.0mg, 0.074 mmol) and N,N′-dimethylethane-1,2-diamine (6.5 mg, 7.8 μL,0.073 mmol) were added and the nitrogen flush was continued for another5 minutes. The tube was sealed and the contents heated at 100° C. for 18hours. After cooling to room temperature, the reaction mixture wasdiluted with 20 mL water and the precipitate collected by filtration.The solid was washed with water, washed with methanol, and then taken upin 6 mL DMSO. Purification by reversed-phase HPLC provided2-(5,6-dimethoxypyridin-3-yl)-6-(5-methylthiophen-2-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 252, 31.6 mg, 0.084 mmol, 23% yield): ESMS (M+H) 368.01; ¹HNMR (300 MHz, DMSO-d₆) δ 8.56 (d, J=2.0 Hz, 1H), 8.30-8.12 (m, 2H), 7.99(d, J=2.0 Hz, 1H), 6.73 (d, J=3.7 Hz, 1H), 6.67 (dd, J=3.7, 1.1 Hz, 1H),5.09 (s, 2H), 3.96 (s, 3H), 3.92 (s, 3H), 2.42 (d, J=0.7 Hz, 3H).

Example 15 Preparation of2-(5,6-dimethoxypyridin-3-yl)-7,7-dimethyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 311)

As shown in step 15-i of Scheme 15, to a solution of2-(5,6-dimethoxypyridin-3-yl)-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 115, 20 mg, 0.047 mmol, prepared by reacting compound 2029with 1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-amine in a manner similar tothat shown in Example 8) in DMF (500 μL) was added iodomethane (17 mg,0.119 mmol) followed by NaH (6 mg, 60% w/w in mineral oil). The reactionwas stirred at room temperature for 2 hours, quenched with a saturatedaqueous solution of NaHCO₃ (1 mL) and extracted with DCM (3×2 mL). Theorganics were concentrated and the crude residue was purified bypreparative silica gel thin layer chromatography (100% EtOAc) to provide2-(5,6-dimethoxypyridin-3-yl)-7,7-dimethyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 346, 11.2 mg, 50% yield) as a white solid::ESMS (M+H) 447.87;¹H NMR (300 MHz, CDCl₃) δ 8.45 (d, J=1.9 Hz, 1H), 8.24-8.08 (m, 2H),7.84 (dd, J=19.0, 4.1 Hz, 3H), 4.75 (q, J=8.3 Hz, 2H), 4.08 (t, J=17.3Hz, 6H), 1.72 (s, 6H).

Example 16 Preparation of(S)-2-(6-ethoxy-5-methoxypyridin-3-yl)-7-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 354) and(R)-2-(6-ethoxy-5-methoxypyridin-3-yl)-7-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 355)

As shown in step 16-i of Scheme 16, to a solution of2,2,2-trifluoroethanol (26.54 g, 19.33 mL, 265 mmol) and pyridine (20.99g, 21.46 mL, 265 mmol) in DCM (120 mL) at 0° C. was added a solution oftrifluoromethylsulfonyl trifluoromethanesulfonate (74.85 g, 44.6 mL, 265mmol) in DCM (150 mL) via addition funnel over the course of 45 minutes.The reaction mixture was stirred an additional 15 minutes aftercompletion of addition then quenched with water (400 mL). The organicswere washed with water (400 mL), dried over MgSO₄, and filtered toproduce 2,2,2-trifluoroethyl trifluoromethanesulfonate (Compound 2045),which was used as is. As shown in step 16-ii of Scheme 16, the solutionof Compound 2045 was added over the course of 25 min to a solution of4-nitro-1H-pyrazole (25 g, 221.1 mmol) in DMF (200 mL) with K₂CO₃ (61.11g, 442.2 mmol) cooled in an ice-water bath. Once addition was complete,the cooling bath was removed and the reaction mixture stirred at 23° C.for 16 hours. The organics were washed with water (500 mL) and theaqueous wash was extracted with DCM (150 mL). The combined organics weredried over MgSO₄, filtered, and concentrated under reduced pressure. Theresulting DMF-containing concentrate was diluted with 1:1 EtOAc:hexanes(500 mL), washed with water (3×250 mL), brine (200 mL), dried (MgSO₄),filtered, and concentrated under reduced pressure to give4-nitro-1-(2,2,2-trifluoroethyl)pyrazole (Compound 2046, 40.4 g, 207.1mmol, 93.65% yield) as a tan solid: ¹H NMR (CDCl₃, 300 MHz) δ 8.31 (s,1H), 8.17 (s, 1H), 4.79 (q, J=9 Hz, 2H).

As shown in step 16-iii of Scheme 16, to a solution of Compound 2046(40.4 g, 207.1 mmol) in EtOH (600 mL) in a Parr bottle was addedpalladium (10 g, 9.397 mmol) (Pd/C, 10 wt % dry basis, wet, Degussatype). The mixture was placed under 50 p.s.i. of hydrogen gas and shakenat 23° C. for 40 minutes. The reaction mixture was through a Corning0.22 μm PES membrane and the filtrate was concentrated to give1-(2,2,2-trifluoroethyl)pyrazol-4-amine (Compound 2047, 33.94 g, 205.6mmol, 99.24% yield) as a clear reddish oil: ¹H NMR (CDCl₃, 300 MHz) δ7.26 (s, 1H), 7.10 (s, 1H), 4.59 (q, J=9 Hz, 2H), 2.95 (br s, 2H). ESMS(M+H) 165.97.

As shown in step 16-iv of Scheme 16, to Compound 2047 (7.16 g, 43.36mmol) in THF (204.6 mL) at 23° C. was addedfuro[3,4-b]pyridine-5,7-dione (6.465 g, 43.36 mmol) followed by theaddition of DMAP (52.97 mg, 0.4336 mmol). The reaction mixture wasstirred at 50° C. After 3 hours, acetic anhydride (8.853 g, 8.182 mL,86.72 mmol) was added and the reaction mixture heated at 70° C. foranother 1.5 hours. After cooling, the reaction mixture was concentratedand the residue partitioned between DCM and saturated aqueous NaHCO₃(100 mL each). The aqueous layer was extracted with DCM (50 mL) and thecombined organics were washed with saturated aqueous NaHCO₃ (100 mL),dried (MgSO₄), filtered, and concentrated under reduced pressure. Theresidue was recrystallized from hot EtOAc/hexanes to give6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[3,4-b]pyridine-5,7(6H)-dione(Compound 2048, 6.07 g, 20.49 mmol, 47.27%) as yellow needles: ESMS(M+H) 297.23; ¹H NMR (CDCl₃, 300 MHz) δ 9.05 (m, 1H), 8.32 (s, 1H), 8.31(s, 1H), 8.26 (m, 1H), 7.70 (m, 1H), 4.79 (q, J=9 Hz, 2H).

As shown in step 16-v of Scheme 16, to Compound 2048 (5.69 g, 19.21mmol) in THF (500 mL) at −78° C. under an atmosphere of nitrogen wasslowly added methyl magnesium bromide (16.95 g, 16.46 mL of 1.4 Msolution in 1:3 THF:toluene, 23.05 mmol). After stirring for 1 hour at−78° C. the reaction was warmed to 0° C. and stirred an additional 1hour. The reaction was quenched with saturated aqueous NH₄Cl (100 mL).After stirring for 15 minutes, the mixture was partially concentratedunder reduce pressure and partitioned between water (150 mL) and EtOAc(200 mL). The organics were washed with brine (150 mL), dried (MgSO₄),filtered, and concentrated to give7-hydroxy-7-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2049, 5.77 g, 18.48 mmol, 96.2% yield) as a yellow solid: ESMS(M+H) 313.23; ¹H NMR (CDCl₃, 300 MHz) δ 8.73 (m, 1H), 8.10 (s, 1H), 8.01(s, 1H), 8.00 (m, 1H), 7.40 (m, 1H), 4.73 (q, J=9 Hz, 2H), 1.84 (s, 3H).

As shown in step 16-yl of Scheme 16, to Compound 2049 (5.77 g, 18.48mmol) in DCM (170 mL) at 23° C. was added Et₃N (7.48 g, 10.30 mL, 73.9mmol) followed by methanesulfonyl chloride (MsCl, 3.18 g, 2.15 mL, 27.7mmol). After stirring for 20 minutes, EtOH (6 mL) was added and stirringcontinued for 10 minutes in order to quench any excess MsCl. Thereaction mixture was partitioned between saturated aqueous NaHCO₃ (300mL) and DCM (50 mL). The aqueous layer was extracted with DCM (150 mL)and the combined organics were dried (MgSO₄), filtered, and combinedwith EtOH (200 mL). The resulting solution (containing7-methylene-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one,Compound 2050) was concentrated under reduced pressure to approximately100 mL and diluted with additional EtOH (250 mL). Pd/C (10 wt % drybasis, wet, Degussa type, 2.85 g) was added and the reaction mixturestirred under an atmosphere of hydrogen for 1 hour as shown in step16-vii of Scheme 16. Analysis showed incomplete conversion of startingmaterial to product so the mixture was filtered, treated with freshcatalyst (3.0 g), then stirred under an atmosphere of hydrogen for 90minutes at 23° C. The catalyst was removed by filtration and theresulting solution concentrated under reduce pressure to give7-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2051, 5.532 g, 18.67 mmol, 100% yield): ESMS (M+H) 297.23; ¹HNMR (CDCl₃, 300 MHz) δ 8.80 (m, 1H), 8.28 (s, 1H), 8.19 (m, 1H), 7.76(s, 1H), 7.47 (m, 1H), 4.98 (q, J=6 Hz, 1H), 4.76 (q, J=9 Hz, 2H), 1.70(d, J=6 Hz, 3H). NMR analysis show a minor amount of over-reducedmaterial but the crude product was used as is in subsequent reactions.

As shown in step 16-viii of Scheme 16, to a solution of Compound 2051(5.532 g, 18.67 mmol) in CHCl₃ (58.20 mL) was added m-CPBA (6.903 g,28.00 mmol). The reaction mixture was stirred at 23° C. for 2 days. Thereaction mixture was partitioned between saturated aqueous NaHCO₃ andDCM (100 mL each) and the aqueous layer extracted with DCM (100 mL). Thecombined organics were dried (MgSO₄), filtered, and concentrated underreduced pressure. The residue was purified by medium pressure silica gelchromatography (0-15% MeOH in DCM) to give7-methyl-5-oxo-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridine1-oxide (Compound 2052, 3.47 g, 11.11 mmol, 59.5% yield) as a whitesolid: ESMS (M+H) 313.23; ¹H NMR (CD₃OD, 300 MHz) δ 8.50 (dd, J=3, 6 Hz,1H), 8.30 (s, 1H), 7.97 (s, 1H), 7.94 (d, J=9 Hz, 1H), 7.69 (t, J=6 Hz,1H), 5.36 (q, J=6 Hz, 1H), 5.00 (q, J=9 Hz, 2H), 1.74 (d, J=6 Hz, 3H).

As shown in step 16-ix of Scheme 16, to Compound 2052 (3.47 g, 11.11mmol) in CHCl₃ (10 mL) at 85° C. was added POCl₃ (17.04 g, 10.36 mL, 111mmol). After 2.5 hours at 85° C., the reaction mixture was treated withtoluene (30 mL) and then concentrated to give a dark purple glassy oil,which was partitioned between DCM and saturated aqueous NaHCO₃ (300 mLeach). An insoluble dark material was observed. The aqueous layer wasextracted with DCM (3×150 mL) and the combined organics dried (MgSO₄),filtered, and concentrated under reduced pressure. The residue waspurified by medium pressure silica gel chromatography (0-80% EtOAc inhexanes) to give2-chloro-7-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2053, 1.20 g, 3.64 mmol, 32.8% yield) as a tan solid: ESMS(M+H) 331.19; ¹H NMR (CDCl₃, 300 MHz) δ 8.26 (s, 1H), 8.13 (d, J=9 Hz,1H), 7.74 (s, 1H), 7.50 (d, J=9 Hz, 1H), 4.95 (q, J=6 Hz, 1H), 4.75 (q,J=9 Hz, 2H), 1.70 (d, J=6 Hz, 3H).

As shown in step 16-x of Scheme 16, to Compound 2053 (366 mg, 1.107mmol),2-ethoxy-3-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(371 mg, 1.328 mmol), Na₂CO₃ (352 mg, 3.32 mmol), and Pd(PPh₃)₄ (64 mg,0.055 mmol) was added DMF (12 mL) followed by water (3 mL). The reactionvessel was evacuated, placed under an atmosphere of hydrogen, and warmedto 100° C. (sand bath). After 18 hours, the reaction mixture waspartitioned between EtOAc and water (100 mL each). The organics werewashed with water (2×80 mL), brine (80 mL) dried (MgSO₄), filtered, andconcentrated under reduced pressure. The residue was dissolved in hotEtOAc (20 mL) then treated with hexanes (20 mL). After standing at 23°C. for 2.5 h the resulting precipitate was collected by filtration anddried in vacuo to give a mixture of(S)-2-(6-ethoxy-5-methoxypyridin-3-yl)-7-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 354) and(R)-2-(6-ethoxy-5-methoxypyridin-3-yl)-7-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 355) (347 mg, 0.7583 mmol, 68.50%) as off-white needles: ESMS(M+H) 448.39; ¹H NMR (CDCl₃, 300 MHz) δ 8.44 (d, J=3 Hz, 1H), 8.31 (s,1H), 8.22 (d, J=9 Hz, 1H), 7.93 (d, J=3 Hz, 1H), 7.86 (d, J=9 Hz, 1H),7.77 (s, 1H), 5.03 (q, J=6 Hz, 1H), 4.77 (q, J=9 Hz, 2H), 4.61 (q, J=6Hz, 2H), 4.04 (s, 3H), 1.76 (d, J=6 Hz, 3H), 1.52 (t, J=6 Hz, 3H). Thesetwo compounds were separated by supercritical fluid chromatography on aWhelk-O-1® (Regis Technologies, Inc.) column using DMF as modifier togive the individual enantiomers.

Example 17 Preparation of2-(5,6-dimethoxypyridin-3-yl)-7-methoxy-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 256)

As shown in step 17-i of Scheme 17, to6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[3,4-b]pyridine-5,7(6H)-dione(Compound 2048, 2.32 g, 7.832 mmol) in AcOH (30 mL) at 23° C. was addedZn (2.561 g, 39.16 mmol). After stirring for 20 minutes at 23° C., thereaction mixture was filtered through a glass frit, and the filtrate wasconcentrated. The residue was dissolved/suspended in hot EtOH (40 mL).The resulting mixture was cooled, treated with Et₂O (50 mL). Theresulting precipitate was collected by filtration and the mother liquorwas concentrated under reduced pressure and the resulting solidrecrystallized from hot EtOH (20 mL) and Et₂O to give additional7-hydroxy-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2055, 1.61 g total) as a yellow solid: ESMS (M+H) 299.26; ¹HNMR (DMSO-d₆, 300 MHz) δ 8.83 (dd, J=3, 6 Hz, 1H), 8.32 (s, 1H), 8.15(dd, J=3, 9 Hz, 1H), 7.94 (s, 1H), 7.61 (dd, J=6, 9 Hz, 1H), 7.13 (d,J=9 Hz, 1H), 6.21 (d, J=9 Hz, 1H), 5.20 (q, J=9 Hz, 2H).

As shown in step 17-ii of Scheme 17, to a solution/suspension ofCompound 2055 (1.16 g, 3.890 mmol) in DCM (20 mL) and THF (10 mL) at 23°C. was added TEA (1.58 g, 2.17 mL, 15.56 mmol) followed by MsCl (668 mg,452 μL, 5.84 mmol). The starting material went into solution over thecourse of 10 minutes. After 1 hour, methanol (10 mL) was added. Afterstirring an additional 2 hours, the mixture was concentrated underreduced pressure. The residue was partitioned between DCM and saturatedaqueous NaHCO₃ (100 mL each) and the aqueous layer extracted with DCM(50 mL). The combined organics were dried (MgSO₄), filtered, andconcentrated under reduced pressure. The residue was purified by mediumpressure silica gel chromatography (0-100% EtOAc in hexanes) to give7-methoxy-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2056, 0.82 g, 2.63 mmol, 67.5% yield) as a white solid: ESMS(M+H) 313.29; ¹H NMR (CDCl₃, 300 MHz) δ 8.89 (dd, J=3, 6 Hz, 1H), 8.26(s, 1H), 8.19 (dd, J=3, 9 Hz, 1H), 8.05 (s, 1H), 7.54 (dd, J=6, 9 Hz,1H), 6.15 (s, 1H), 4.76 (q, J=9 Hz, 2H), 3.12 (s, 3H).

As shown in step 17-iii of Scheme 17, to a solution of Compound 2056(0.82 g, 2.63 mmol) in CHCl₃ (10 mL) was added mCPBA (777 mg, 3.15mmol). The reaction was stirred at 23° C. for 5 minutes, warmed to 59°C. (sand bath), stirred at 59° C. for 24 hours, then at 23° C. for anadditional 3 days. Purification by medium pressure silica gelchromatography (0-15% MeOH in DCM) gave7-methoxy-5-oxo-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridine1-oxide (Compound 2057, 628 mg, 1.91 mmol, 73% yield) as a white solid:ESMS (M+H) 329.23; ¹H NMR (CDCl₃, 300 MHz) δ 8.49 (dd, J=2, 6 Hz, 1H),8.31 (s, 1H), 7.89 (s, 1H), 7.71 (m, 2H), 6.52 (s, 1H), 5.21 (q, J=9 Hz,2H), 3.18 (s, 3H).

As shown in step 17-iv of Scheme 17, to Compound 2057 (620 mg, 1.89mmol) was added CHCl₃ (3.5 mL) followed by the addition of POCl₃ (5.79g, 3.52 mL, 37.8 mmol). The reaction mixture was heated to 90° C. (sandbath). After 1.8 hours, toluene (10 mL) was added then the solution wasconcentrated under reduced pressure to remove excess POCl₃. The residuewas partitioned between DCM and saturated aqueous NaHCO₃ (100 mL each)and the aqueous layer was extracted with DCM (50 mL). The combinedorganics were dried (MgSO₄), filtered, concentrated under reducedpressure, and the residue purified by medium pressure silica gelchromatography (0-65% EtOAc in hexanes) to give2-chloro-7-methoxy-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2058, 275 mg, 0.79 mmol, 42% yield) as a clear oil: ESMS (M+H)347.18; ¹H NMR (CDCl₃, 300 MHz) δ 8.23 (s, 1H), 8.13 (d, J=9 Hz, 1H),8.02 (s, 1H), 7.57 (d, J=9 Hz, 1H), 6.08 (s, 1H), 4.76 (q, J=9 Hz, 2H),3.18 (s, 3H).

As shown in step 17-v of Scheme 17, to Compound 2058 (293 mg, 0.85mmol),2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(269 mg, 1.01 mmol), Na₂CO₃ (179 mg, 1.69 mmol), and Pd(PPh₃)₄ (98 mg,0.085 mmol) was added DMF (10 mL), followed by the addition of water (2mL). The reaction vessel was evacuated, placed under an atmosphere ofnitrogen, then warmed to 110° C. (sand bath). After 16 hours thereaction mixture was partitioned between EtOAc and water (100 mL each).The organics were washed with water (70 mL), brine (70 mL), dried(MgSO₄), filtered, and concentrated under reduced pressure. The residuewas dissolved/suspended in EtOAc (7 mL) and heated with swirling in awater bath at 50° C. for 40 minutes. The resulting mixture was treatedwith hexanes (5 mL) and swirled an additional 5 minutes at 50° C. Aftercooling to 23° C., the resulting solid was collected by filtration andwashed with 1:1 (EtOAc:hexanes, 5 mL) to give2-(5,6-dimethoxypyridin-3-yl)-7-methoxy-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 256) as a mixture of enantiomers: ESMS (M+H) 450.44; ¹H NMR(CDCl₃, 300 MHz) δ 8.57 (d, J=3 Hz, 1H), 8.35 (s, 1H), 8.27 (s, 2H),7.99 (d, J=3 Hz, 1H), 7.93 (s, 1H), 6.40 (s, 1H), 5.22 (q, J=9 Hz, 2H),3.96 (s, 3H), 3.93 (s, 3H), 3.13 (s, 3H).

Example 18 Preparation of2-(5,6-dimethoxypyridin-3-yl)-7-(2-methoxyethoxy)-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 336)

As shown in step 18-i of Scheme 18, to7-hydroxy-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2055, 4.35 g, 14.6 mmol) in CHCl₃ (80 mL) and MeOH (40 mL) wasadded mCPBA (5.39 g, 21.9 mmol). After stirring for 24 hours, additionalmCPBA (1.26 g, 7.30 mmol) was added. After stirring an additional 16hours, the resulting precipitate was collected by filtration and washedwith DCM (10 mL) and Et₂O (20 mL) to give7-hydroxy-5-oxo-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridine1-oxide (Compound 2059, 2.49 g) as a white solid: ESMS (M+H) 315.25; ¹HNMR (DMSO-d₆, 300 MHz) δ 8.44 (d, J=6 Hz, 1H), 8.31 (s, 1H), 7.89 (s,1H), 7.65 (m, 2H), 7.43 (d, J=9 Hz, 1H), 6.41 (d, J=9 Hz, 1H), 5.20 (q,J=9 Hz, 2H).

As shown in step 18-ii of Scheme 18, to Compound 2059 (312 mg, 0.99mmol) in MeCN (10 mL) was added K₂CO₃ (686 mg, 4.96 mmol) followed bythe addition of POCl₃ (761 mg, 463 μL, 4.96 mmol). The reaction mixturewas refluxed under an atmosphere of nitrogen for 24 hours and thenfiltered through a glass frit. The filtrate was concentrated underreduced pressure then partitioned between DCM (60 mL), water (10 mL),and saturated aqueous NaHCO₃ (50 mL). The aqueous layer was extractedwith DCM (50 mL) and the combined organics were dried (MgSO₄), filtered,and concentrated under reduced pressure. The residue was purified bymedium pressure silica gel chromatography (0-60% EtOAc in hexanes) togive2,7-dichloro-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2060, 150 mg, 0.43 mmol, 43%) as a white solid: ¹H NMR (CDCl₃,300 MHz) δ 8.14 (s, 1H), 8.07 (d, J=9 Hz, 1H), 7.89 (s, 1H), 7.51 (d,J=9 Hz, 1H), 6.50 (s, 1H), 4.69 (q, J=9 Hz, 2H).

As shown in step 18-ii of Scheme 18, to Compound 2060 (180 mg, 0.5127mmol) in DMF (6 mL) was added2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2021, 163 mg, 0.62 mmol), Na₂CO₃ (217 mg, 2.05 mmol),Pd(PPh₃)₄ (30 mg, 0.026 mmol), and 2-methoxyethanol (1.5 mL, 19.0 mmol).The reaction vessel was evacuated, placed under an atmosphere ofnitrogen, then warmed to 100° C. (sand bath). After 16 hours, thereaction mixture was partitioned between EtOAc and water (100 mL each).The organics were concentrated under reduced pressure and the residuedissolved in DMSO (5 mL) and purified by reversed-phase HPLC (10-90%aqueous MeCN with 0.1% TFA buffer) to give2-(5,6-dimethoxypyridin-3-yl)-7-(2-methoxyethoxy)-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 336, 55 mg) as a white lyophilizate: ESMS (M+H) 494.39; ¹H NMR(CD₃OD, 300 MHz) δ 8.50 (d, J=3 Hz, 1H), 8.33 (s, 1H), 8.22 (d, J=9 Hz,1H), 8.13 (d, J=9 Hz, 1H), 8.08 (d, J=3 Hz, 1H), 8.00 (s, 1H), 6.29 (s,1H), 4.99 (q, J=9 Hz, 2H), 4.03 (s, 3H), 3.97 (s, 3H), 3.66 (m, 1H),3.53 (m, 3H), 3.29 (s, 3H).

Example 19 Preparation of methyl2-(2-(5,6-dimethoxypyridin-3-yl)-5-oxo-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl)acetate(Compound 307)

As shown in step 19-i of Scheme 19, to7-hydroxy-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 2055, 1.75 g, 5.87 mmol) and methyl2-triphenylphosphoranylideneacetate (2.06 g, 6.16 mmol) was addedtoluene (23 mL) and THF (12 mL). The reaction mixture was heated toreflux and held there for 2.5 hours. After cooling, the reaction mixturewas concentrated under reduced pressure and the residue purified bymedium pressure silica gel chromatography (0-7.5% EtOH in DCM) to givemethyl2-(5-oxo-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl)acetate(Compound 2061, 2.25 g, 6.35 mmol) as a clear oil: ESMS (M+H) 355.29.The product contained a small amount of triphenylphosphine oxide but wasused in subsequent reactions as is.

As shown in step 19-ii of Scheme 19, to Compound 2061 (2.08 g, 5.89mmol) in CHCl₃ (32 mL) was added mCPBA (2.17 g, 8.80 mmol) and thereaction mixture refluxed for 2 hours, cooled, and concentrated underreduced pressure. The resulting residue was purified by medium pressuresilica gel chromatography (0-12% EtOH in DCM) to give7-(2-methoxy-2-oxoethyl)-5-oxo-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridine1-oxide (Compound 2062, 1.31 g, 3.55 mmol, 60% yield) as a white solid:ESMS (M+H) 371.35; ¹H NMR (DMSO-d₆, 300 MHz) δ 8.48 (d, J=6 Hz, 1H),8.30 (s, 1H), 7.91 (s, 1H), 7.75 (d, J=6 Hz, 1H), 7.63 (t, J=3 Hz, 1H),5.63 (m, 1H), 5.21 (q, J=9 Hz, 2H), 3.56 (dd, J=3, 15 Hz, 1H), 3.36 (s,3H), 3.16 (dd, J=6, 15 Hz, 1H).

As shown in step 19-iii of Scheme 19, to Compound 2062 (1.06 g, 2.86mmol) was added POCl₃ (13.17 g, 8.00 mL, 85.9 mmol). The reactionmixture was heated at 90° C. (sand bath) for 2.5 hours, followed by theremoval of POCl₃ under reduced pressure. The residue was partitionedbetween saturated aqueous NaHCO₃ and DCM (100 mL each) and the aqueouslayer was extracted with DCM (50 mL). The combined organics were dried(MgSO₄), filtered, and concentrated under reduced pressure. The residuewas purified by medium pressure silica gel chromatography (0-100% EtOAcin hexanesto give methyl2-(2-chloro-5-oxo-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl)acetate(Compound 2063, 378 mg, 0.97 mmol, 34% yield) as a white solid: ESMS(M+H) 389.33; ¹H NMR (CDCl₃, 300 MHz) δ 8.13 (d, J=9 Hz, 1H), 8.12 (s,1H), 7.75 (s, 1H), 7.51 (d, J=9 Hz, 1H), 5.30 (m, 1H), 4.76 (q, J=9 Hz,2H), 3.62 (s, 3H), 3.10 (m, 2H).

As shown in step 19-iv of Scheme 19, to Compound 2063 (375 mg, 0.965mmol),2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2021, 307 mg, 1.16 mmol), Na₂CO₃ (205 mg, 1.93 mmol),Pd(PPh₃)₄ (112 mg, 0.0965 mmol) was added DMF (12 mL) followed by theaddition of water (2.5 mL). The reaction vessel was evacuated, placedunder an atmosphere of nitrogen, then warmed to 110° C. (sand bath).After 18 hours, the reaction mixture was cooled and partitioned betweenEtOAc and water (100 mL each). The organics were washed with brine (50mL), dried (MgSO₄), filtered and concentrated under reduced pressure.The residue was suspended in hot EtOAc (20 mL) and swirled for 45 min at60° C. to give a uniform suspension, which was allowed to stand at 23°C. for 24 hours. The resulting solid was collected by filtration,dissolved in warm DMSO (50 mL), and filtered through a 0.45 micron PTFEmembrane (syringe filter). The filtrate was treated with water (5 mL)and the resulting precipitate collected by filtration, washed with water(10 mL), suspended in warm MeCN (5 mL) and treated with water (5 mL).The resulting suspension was frozen and lyophilized to give methyl2-(2-(5,6-dimethoxypyridin-3-yl)-5-oxo-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl)acetate(Compound 307, 195 mg, 0.38 mmol, 39% yield) as a white solid: ESMS(M+H) 491.86; ¹H NMR (DMSO-d₆, 300 MHz) δ 8.56 (d, J=3 Hz, 1H), 8.30 (s,1H), 8.23 (d, J=9 Hz, 1H), 8.19 (d, J=9 Hz, 1H), 7.99 (d, J=3 Hz, 1H),7.89 (s, 1H), 5.51 (m, 1H), 5.21 (q, J=9 Hz, 2H), 3.96 (s, 3H), 3.92 (s,3H), 3.46 (s, 3H), 3.26 (dd, J=3, 15 Hz, 1H), 3.03 (dd, J=6, Hz, 1H).

Example 20 Preparation of6-(1-(2-(1H-pyrazol-1-yl)ethyl)-1H-pyrazol-4-yl)-2-(5,6-dimethoxypyridin-3-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 171)

As shown in step 20-i of Scheme 20,2-(5,6-dimethoxypyridin-3-yl)-6-(1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one[Compound 70, 100 mg, 0.2964 mmol, prepared from2-chloro-6-(1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-oneand2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2021) in a manner similar to the preparation of compound 135as shown in Example 12], and cesium carbonate (193 mg, 0.593 mmol), wereweighed into a conical microwave vial equipped with a stir bar. DMF(1.05 mL) was added followed by the addition of1-(2-chloroethyl)pyrazole (77 mg, 0.593 mmols). The vial was sealed andheated at 120° C. for 15 minutes. Water (3 mL) was added and theresulting precipitate collected by filtration and washed with 5 mL ofwater. The filtrate was concentrated under reduced pressure. Each of thecollected solid and the residue from concentration of the filtrate wasdiluted with DMSO until solubilized and purified by reversed-phase HPLCto give6-(1-(2-(1H-pyrazol-1-yl)ethyl)-1H-pyrazol-4-yl)-2-(5,6-dimethoxypyridin-3-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 171, 22 mg, 0.05 mmol, 17% yield): ESMS (M+H) 432.0; ¹H NMR(DMSO-d₆, 300 MHz) δ 8.47 (d, J=2.0 Hz, 1H), 8.10 (s, 2H), 7.95-7.87 (m,2H), 7.75 (s, 1H), 7.45 (d, J=2.2 Hz, 1H), 7.40 (d, J=1.3 Hz, 1H), 6.12(t, J=2.0 Hz, 1H), 4.84 (s, 2H), 4.50 (s, 4H), 3.88 (s, 3H), 3.85 (s,3H).

Example 21 Preparation of6-(5,6-dimethoxypyridin-3-yl)-4-methyl-2-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-1H-pyrrolo[3,4-c]pyridin-3(2H)-one(Compound 243)

As shown in step 21-i of Scheme 21, to a round-bottomed flask containing1-(2,2,2-trifluoroethyl)pyrazol-4-amine (2.01 g, 12.2 mmol) andpotassium carbonate (3.364 g, 24.34 mmol) under an atmosphere ofnitrogen was added DMF (15 mL), followed by the addition of3-bromoprop-1-yne (1.45 g, 1.09 mL, 12.2 mmol). The reaction mixture wasstirred at RT for 18 hours. Water and EtOAc were added and the aqueouslayer extracted with EtOAc. The combined organics were washed withbrine, water, dried (sodium sulfate), filtered, and concentrated underreduced pressure. Purification by silica gel chromatography (petroleumether:EtOAc, 1:1) gaveN-(prop-2-ynyl)-1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-amine (Compound2064, 1.19 g, 48% yield) as an orange solid: ¹H NMR (CDCl₃, 300 MHz) δ7.32 (1H, s), 7.16 (1H, s), 4.62 (2H, q), 3.81 (2H, dd), 3.20 (1H, t),2.27 (1H, t).

As shown in step 21-ii of Scheme 21, to a solution of Compound 2064(1.19 g, 5.88 mmol) in DCM (20 mL) was added DIEA (2.28 g, 3.07 mL, 17.6mmol), but-2-ynoic acid (544 mg, 6.47 mmol) and DMAP (36 mg, 0.29 mmol).The reaction mixture was cooled in ice-bath, EDCI (1.05 g, 6.76 mmol)was added, and the cold bath removed after 3 minutes. After stirring thereaction mixture for 18 hours at RT, water and DCM were added and thelayers separated. The aqueous layer was extracted with DCM and thecombined organics were washed with brine, water, dried (sodium sulfate),filtered and concentrated under reduced pressure. Purification by silicagel chromatography (petroleum ether:EtOAc, 1:1) gaveN-(prop-2-ynyl)-N-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)but-2-ynamide(Compound 2065, 650 mg, 41% yield) as a white solid: ¹H NMR (CDCl₃, 300MHz) δ 8.18 (1H, s), 7.77 (1H, s), 7.71 (1H, s), 7.69 (1H, s), 4.78 (2H,d), 4.76-4.63 (2H, m), 4.46 (2H, d), 2.40 (1H, t), 2.27 (1H, t), 2.13(2H, s), 1.83 (2H, s).

As shown in step 21-iii of Scheme 21, to a solution of ethylN-(oxomethylene)carbamate (385 mg, 345 μL, 3.34 mmol) and Cp*RuCl(cod)(21 mg, 0.056 mmol) in dry 1,2-dichloroethane (3 mL) was added asolution of Compound 2065 (303 mg, 1.11 mmol) in 1,2-dichloroethane (6mL) over 25 min under nitrogen at rt. The reaction mixture was heated at65° C. for 1 hr then concentrated under reduced pressure. Purificationby silica gel chromatography (petroleum ether:EtOAc, 1/1 gradient to0/1) gave ethyl4-methyl-3,6-dioxo-2-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-5(6H)-carboxylate(Compound 2066, 179 mg, 41% yield) as a white solid: ¹H NMR (DMSO-d₆,300 MHz) δ 8.28 (1H, s), 7.84 (1H, d), 6.52 (1H, s), 5.18 (2H, q), 4.75(2H, s), 4.51 (2H, q), 2.67 (2H, s), 1.36 (2H, t).

As shown in step 21-iv of Scheme 21, to a solution of Compound 2066 (614mg, 1.58 mmol) in THF (10 mL) was added HCl (6 M, 10 mL) at rt. Thereaction mixture was heated at reflux overnight and then concentratedunder reduced pressure. To the residue was added phosphorus oxychloride(15 mL, 161 mmol) and the reaction mixture was heated at 95° C. for 3hours under an atmosphere of nitrogen. After cooling and concentratingthe mixture under reduced pressure, ice was added followed by theaddition of EtOAc 30 minutes later. The aqueous layer was extracted withEtOAc and the combined organics washed with brine, dried (sodiumsulfate), filtered, and concentrated under reduced pressure.Purification by silica gel chromatography (petroleum ether:EtOAc, 1:1)gave6-chloro-4-methyl-2-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-1H-pyrrolo[3,4-c]pyridin-3(2H)-one(Compound 2067, 356 mg, 68%) as a white solid: ESMS (M+H) 330.90.

As shown in step 21-iv of Scheme 21, Compound 2067 (129 mg, 0.390 mmol)and Compound 2021 (134 mg, 0.51 mmol) were taken up in DMF and sodiumcarbonate (1 M, 0.780 mmol) and nitrogen passed through the solution for30 minutes. Tetrakistriphenylphosphine palladium(0) (23 mg, 0.020 mmol)was added and the reaction mixture flushed with nitrogen for anadditional 5 min, then heated at 110° C. overnight. After cooling thereaction to room temperature, EtOAc and water were added. The aqueouslayer was extracted with EtOAc and the combined organics were dried(sodium sulfate), filtered through diatomaceous earth, and concentratedunder reduced pressure. Purification by silica gel chromatography(EtOAc) gave6-(5,6-dimethoxypyridin-3-yl)-4-methyl-2-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-1H-pyrrolo[3,4-c]pyridin-3(2H)-one(Compound 243, 137 mg; 77% yield) as a white solid.

Example 22 Preparation of2-(5,6-dimethoxypyridin-3-yl)-7-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Compound 254)

As shown in step 22-i of Scheme 22, a 1 L round bottom flask fitted witha condenser was charged with 2,6-dichloropyridine-3-carboxylic acid(10.0 g, 52.1 mmol),2,3-dimethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2021, 13.81 g, 52.1 mmol), Pd(PPh₃)₄ (3.01 g, 2.60 mmol),Na₂CO₃ (16.56 g, 156 mmol), dioxane (250 mL) and water (100 mL). Theflask was evacuated for 1 minute and the mixture placed under anatmosphere of N₂. The mixture was heated at 110° C. for 16 hours, afterwhich time a precipitate formed. The reaction mixture was cooled andtransferred to a separatory funnel. Na₂CO₃ (200 mL 10 wt % aqueous) wasadded, followed by water (100 mL) and EtOAc (500 mL). Theprecipitate/emulsion persisted, mostly localized in the aqueous layer.The aqueous layer was separated, washed with EtOAc (300 mL), and thencarefully acidified with concentrated HCl(˜50 mL) to a pH of 2. Theresulting precipitate was collected via filtration and washed with water(50 mL). The wet solid was transferred to a 1 L flask with aid of EtOH(200 mL) then evaporated to dryness. The solid residue wasdissolved/suspended in EtOAc (120 mL) then treated with hexanes (120mL). The resulting solid was collected via filtration, washed withhexanes (50 mL), and dried under reduced pressure to give2-chloro-6-(5,6-dimethoxy-3-pyridyl)pyridine-3-carboxylic acid (Compound2068, 11.99 g, 78% yield) as a an off-white solid: ESMS (M+H) 295.27; ¹HNMR (DMSO-d₆, 300 MHz) δ 13.72 (s, 1 H), 8.49 (d, J=1.98, 1H), 8.30 (d,J=8.08, 1H), 8.15 (d, J=8.11, 1H), 7.88 (d, J=1.98, 1H), 3.95 (s, 3H),3.91 (s, 3H).

As shown in step 22-ii of Scheme 22, to a solution of1-(2,2,2-trifluoroethyl)pyrazol-4-amine (Compound 2047, 5.78 g, 35.0mmol), 2-chloro-6-(5,6-dimethoxy-3-pyridyl)pyridine-3-carboxylic acid(Compound 2068, 9.38 g, 31.8 mmol) and HBTU (13.28 g, 35.0 mmol) in DMF(150 mL) at 23° C. was added DIEA (12.35 g, 16.64 mL, 95.52 mmol). Thereaction mixture was stirred for 2 h and then partitioned between EtOAc(400 mL) and water (400 mL). The organic layer was separated, washedwith water (400 mL), 10% aqueous Na₂CO₃ (300 mL), brine (300 mL),combined brine and 2 N HCl (300 mL, 20 mL), and brine (300 mL). Theorganic layer was then diluted with EtOAc (150 mL) and EtOH (70 mL) andwarmed to 75° C. with swirling to give a clear solution. The solutionwas treated with MgSO₄ and filtered while still warm. Afterconcentration under reduced pressure, the residue wasdissolved/suspended in EtOAc (200 mL) and spun at 80° C. for 1 h to givea uniform suspension. Hexanes (200 mL) were added and the resultingsuspension was left standing at 23° C. for 14 h. The precipitate wascollected via filtration, washed with hexanes (100 mL) to provide2-chloro-6-(5,6-dimethoxy-3-pyridyl)-N-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]pyridine-3-carboxamide(Compound 2069, 11.32 g, 80% yield) as a white solid after drying invacuo: ESMS (M+H) 442.50; ¹H NMR (DMSO-d₆, 300 MHz) δ 10.86 (s, 1H),8.49 (d, J=1.89, 1H), 8.25 (s, 1H), 8.16 (m, 2H), 7.88 (d, J=1.89, 1H),7.66 (s, 1H), 5.16 (q, J=9.10, 2H), 3.95 (s, 3H), 3.92 (s, 3H).

As shown in step 22-iii of Scheme 22, a 250 mL Parr vessel was chargedwith2-chloro-6-(5,6-dimethoxy-3-pyridyl)-N-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]pyridine-3-carboxamide(Compound 2069, 5.00 g, 11.32 mmol), PdCl₂(CH₃CN)₂ (147 mg, 0.566 mmol),and dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (378 mg,0.792 mmol). Nitrogen was bubbled through mixture for 3 min thentriethylamine (5.727 g, 7.888 mL, 56.60 mmol) was added followed byethynyl(trimethyl)silane (3.34 g, 4.80 mL, 34.0 mmol) were added. Thevessel was sealed and warmed to 100° C. After 14 hours, the reactionmixture was cooled to 23° C. and partitioned between EtOAc (300 mL) andwater (300 mL). The organic layer was separated, washed with water (300mL), saturated aqueous NaHCO₃ (200 mL), brine (300 mL), dried (MgSO₄),filtered, and concentrated under reduced pressure. The residue waspurified by medium pressure silica gel chromatography: 0-100% EtOAc inhexanes to provide 3.7 g of a tan solid. The solid was dissolved inEtOAc (15 mL, hot, 70° C.) then treated with hexanes (30 mL). Theresulting suspension was spun and cooled via an ice-water bath for 40min then the precipitate was collected via filtration, washed withhexanes (30 mL) to give6-(5,6-dimethoxy-3-pyridyl)-N-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]-2-(2-trimethylsilylethynyl)pyridine-3-carboxamide(Compound 2070, 3.14 g, 55%) as a pale yellow solid: ESMS (M+H) 504.63;¹H NMR (CDCl₃, 300 MHz) δ 9.69 (s, 1H), 8.54 (d, J=8.44, 1H), 8.36 (d,J=1.96, 1H), 8.28 (s, 1H), 7.87 (d, J=1.93, 1H), 7.83 (d, J=8.48 Hz,1H), 7.62 (s, 1H), 4.71 (q, J=8.33, 2H), 4.11 (s, 3H), 4.03 (s, 3H).

As shown in step 22-iv of Scheme 22, to a solution of6-(5,6-dimethoxy-3-pyridyl)-N-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]-2-(2-trimethylsilylethynyl)pyridine-3-carboxamide(Compound 2070, 540 mg, 1.07 mmol) in EtOH (18.5 mL) at 23° C. was addeddropwise EtONa (165 μL of a 1.3 M solution in EtOH, 0.214 mmol). After25 min, the resulting slurry was cooled to 0° C. and, after stirring for10 min at 0° C., the slurry was filtered, and the collected solid waswashed with ice-cold EtOH (3×10 mL) to give2-(5,6-dimethoxy-3-pyridyl)-7-methylene-6-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]pyrrolo[3,4-b]pyridin-5-one(Compound 2071, 436 mg, 93%) as a pale yellow solid after drying invacuo: ESMS (M+H) 432.52; ¹H NMR (CDCl₃, 300 MHz) δ 8.38 (d, J=1.96,1H), 8.15 (d, J=8.13, 1H), 7.88 (d, J=1.98, 1H), 7.84 (s, 1H), 7.82 (d,J=8.16 Hz, 1H), 7.74 (s, 1H), 5.86 (d, J=1.89, 1H), 5.11 (d, J=1.89,1H), 4.72 (q, J=8.32, 2H), 4.05 (s, 3H), 3.96 (s, 3H).

As shown in step 22-v of Scheme 22, to a solution of2-(5,6-dimethoxy-3-pyridyl)-7-methylene-6-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]pyrrolo[3,4-b]pyridin-5-one(Compound 2071, 436 mg, 1.01 mmol) in THF (20 mL) was added Pd/C (200mg, 10 wt % dry basis, wet, Degussa type). The reaction vessel wasevacuated and then placed under an atmosphere of H₂ (balloon). Afterstirring for 2.5 h, the reaction mixture was filtered through a pad ofsilica and washed with THF (80 mL). The resulting filtrate wasconcentrated under reduced pressure and the residue treated with EtOAc(6 mL). After heating to reflux to give a uniform suspension, hexanes(10 mL) were added. The resulting suspension was cooled to 0° C.(ice-water bath), held at 0° C. for 5 min, and the precipitate collectedvia filtration to give2-(5,6-dimethoxy-3-pyridyl)-7-methyl-6-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]-7H-pyrrolo[3,4-b]pyridin-5-one(Compound 254, 300 mg, 68%) as a pale tan-colored solid: ESMS (M+H)434.44; ¹H NMR (DMSO-d₆, 300 MHz) δ 8.57 (d, J=3 Hz, 1H), 8.39 (s, 1H),8.23 (d, J=9 Hz, 1H), 8.18 (d, J=9 Hz, 1H), 8.00 (d, J=3 Hz, 1H), 7.97(s, 1H), 5.22 (m, 3H), 3.96 (s, 3H), 3.93 (s, 3H), 1.60 (d, J=6 Hz, 3H).

Example 23 Preparation of2′-(5-methoxypyridin-3-yl)-4′-methyl-6′-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)spiro[cyclopropane-1,7′-pyrrolo[3,4-b]pyridin]-5′(6′H)-one(Compound 651)

As shown in step 23-i of Scheme 23, 1,2-dibromoethane (369.3 mg, 169.4μL, 1.966 mmol) was added to a stirring solution of2-chloro-4-methyl-6-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]-7H-pyrrolo[3,4-b]pyridin-5-one(500 mg, 1.512 mmol, prepared from the reaction of1-(2,2,2-trifluoroethyl)-4-aminopyrazole with Compound 2041) in DMF (12mL) at RT, followed by the addition of NaH (133 mg, 3.326 mmol, 60 wt %dispersion in mineral oil). The reaction mixture was stirred for 30 minat RT, cooled to 0° C. and quenched with sat'd NaHCO₃ (10 mL). Thereaction mixture was extracted with DCM (3×10 mL) and the combinedorganics were dried over Na₂SO₄, filtered, and concentrated underreduced pressure. The crude residue was purified by medium pressuresilica gel chromatography (0-50% EtOAc/hexanes) to provide2′-chloro-4′-methyl-6′-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)spiro[cyclopropane-1,7′-pyrrolo[3,4-b]pyridin]-5′(6′H)-one(Compound 2073, 250 mg, 47% yield): ESMS (M+H) 358.0.

As shown in step 23-ii of Scheme 23, Potassium acetate (20.64 mg, 0.2103mmol) and Pd(PPh₃)₄ (16.20 mg, 0.01402 mmol) were added to a solution ofCompound 2073 (50 mg, 0.1402 mmol) and3-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(Compound 2074, 49.44 mg, 0.2103 mmol) in DMF (383.6 μL) and H₂O (127.9μL). The solution was degassed and then heated to 100° C. in a microwavefor 1 hour. The reaction was concentrated and the residue was purifiedby medium pressure silica gel chromatography (0-100% EtOAc/hexanes) toprovide2′-(5-methoxypyridin-3-yl)-4′-methyl-6′-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)spiro[cyclopropane-1,7′-pyrrolo[3,4-b]pyridin]-5′(6′H)-one(Compound 651, 30 mg, 47% yield) as a white solid: ESMS (M+H) 430.59.

Using the appropriate intermediates, protected if so required, Compounds832, 833, 957, and 966 were also prepared by a similar procedure. Thisprocedure was also used as an alternative to the procedure described inExample 15 for the preparation of Compound 311.

Example 24 Preparation of2-(5-methoxy-3-pyridyl)-4-methyl-641-(2,2,2-trifluoroethyl)pyrazol-4-yl]spiro[pyrrolo[3,4-b]pyridine-7,3′-tetrahydrofuran]-5-one(Compound 972)

As shown in step 24-i of Scheme 24, trimethylsilylcyanide (4.241 g,5.700 mL, 42.75 mmol) was added to a solution of1-(2,2,2-trifluoroethyl)pyrazol-4-amine (7.059 g, 42.75 mmol) andtetrahydrofuran-3-one (3.68 g, 42.75 mmol) in AcOH (45 mL) at 0° C. viasyringe over 30 seconds. The reaction mixture was slowly warmed to 23°C. After stirring for 16 hours, the mixture was added to 1:1 ammoniumhydroxide:ice (200 mL) and extracted with DCM (2×200 mL). The organicswere dried (magnesium sulfate) filtered and concentrated. The residuewas purified by medium pressure silica gel chromatography (0-100% EtOAcin hexanes to provide3-[[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]amino]tetrahydrofuran-3-carbonitrile(Compound 2075, 3.66 g, 14.07 mmol, 32.91% yield) as a brown oil: ESMS(M+H) 261.32; ¹H NMR (CDCl₃, 300 MHz) δ 7.47 (s, 1H), 7.46 (s, 1H), 4.68(q, J=9, 2H), 4.06 (m, 4H), 2.42 (m, 3H).

As shown in step 24-ii of Scheme 24, to a solution of but-2-ynoic acid(833.6 mg, 9.915 mmol) in DCM (12 mL) at 0° C. was added1-chloro-N,N,2-trimethyl-prop-1-en-1-amine (1.325 g, 1.312 mL, 9.915mmol). After stirring for 40 minutes, the reaction mixture was cooled to−78° C. and a solution of3-[[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]amino]tetrahydrofuran-3-carbonitrile(Compound 2075, 1.72 g, 6.610 mmol) and DIEA (2.563 g, 3.454 mL, 19.83mmol) in DCM (12 mL) was added. The reaction mixture was warmed to 0° C.(ice-water bath), and after 1 hour the mixture was warmed to 23° C.After 30 min at 23° C., the reaction mixture was partitioned betweenwater and EtOAc (100 mL each). The organics were separated (insolubleprecipitate is present), washed with water then brine (100 mL each),dried (magnesium sulfate), filtered, and concentrated under reducedpressure. The residue was purified by medium pressure silica gelchromatography (0-80% EtOAc/hexanes) to provideN-(3-cyanotetrahydrofuran-3-yl)-N-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]buta-2,3-dienamide(Compound 2076, 1.48 g) as a yellow oil: ESMS (M+H) 327.20; ¹H NMR(CDCl₃, 300 MHz) δ 7.70 (s, 1H), 7.63 (s, 1H), 5.57 (t, J=6, 1H), 5.19(d, J=6, 2H), 4.77 (q, J=9, 2H), 4.00 (m, 4H), 2.50 (m, 1H), 2.30 (m,1H).

As shown in step 24-iii of Scheme 24, to a solution of ditert-butylpropanedioate (3.579 g, 3.705 mL, 16.55 mmol) in THF (50 mL) at 23° C.was added NaH (496.4 mg, 12.41 mmol). After stirring for 20 minutes, asolution of Compound 2076 (2.70 g, 8.275 mmol) in THF (50 mL) was added.After 1 hour, the reaction was quenched with saturated aqueous ammoniumchloride (100 mL) and partitioned between water and EtOAc (150 mL each).The organics were separated, washed with brine (200 mL) containing 1 NHCl (10 mL aq), washed with brine (150 mL), dried (magnesium sulfate),filtered and concentrated under reduced pressure. The residue waspurified by medium pressure silica gel chromatography (0-100% EtOAc inhexanes) to provide intermediate tert-butyl4′-methyl-2′,5′-dioxo-6′-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-1′,2′,4,5,5′,6′-hexahydro-2H-spiro[furan-3,7′-pyrrolo[3,4-b]pyridine]-3′-carboxylate(2.03 g, 4.334 mmol) as a pale yellow solid: ESMS (M+H) 469.31; ¹H NMR(DMSO-d₆, 300 MHz) δ 12.80 (br s, 1H), 8.16 (s, 1H), 7.80 (s, 1H), 5.17(q, J=9, 2H), 4.18 (d, J=12, 1H), 3.90 (m, 2H), 3.75 (m, 1H), 2.45 (m,1H), 2.44 (s, 3H), 2.33 (m, 1H), 1.52 (s, 9H).

As shown in step 24-iv of Scheme 24, tert-butyl4′-methyl-2′,5′-dioxo-6′-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-1′,2′,4,5,5′,6′-hexahydro-2H-spiro[furan-3,7′-pyrrolo[3,4-b]pyridine]-3′-carboxylate(2.00 g, 4.270 mmol) was taken up in DCM (25 mL) at 23° C. and TFA (25mL) was added. After 30 minutes, the reaction mixture was treated withtoluene (80 mL) and concentrated under reduced pressure to provide4′-methyl-2′,5′-dioxo-6′-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-1′,2′,4,5,5′,6′-hexahydro-2H-spiro[furan-3,7′-pyrrolo[3,4-b]pyridine]-3′-carboxylicacid (Compound 2077, 1.776 g, 4.307 mmol, 52.37% overall yield) as anoff-white solid: ESMS (M+H) 413.32; ¹H NMR (DMSO-d₆, 300 MHz) δ 13.50(br s, 1 H), 8.16 (s, 1H), 7.79 (s, 1H), 5.17 (q, J=9, 2H), 4.15 (d,J=12, 1H), 3.99 (m, 2H), 3.85 (m, 1H), 2.63 (s, 3H), 2.55 (m, 1H), 2.35(m, 1H).

As shown in step 24-v of Scheme 24, to a suspension of Compound 2077(1.77 g, 4.293 mmol) in MeCN (10 mL) was added LiOH dihydrate (270.2 mg,6.440 mmol) followed by water (10 mL). After stirring for 5 minutes, NBS(802.4 mg, 4.508 mmol) was added. After a total of 40 min following theaddition of NBS, the reaction mixture was partitioned between EtOAc andwater (100 mL each). The aqueous layer was treated with 1 N aq HCl (8mL) to give a white precipitate. The resulting suspension wasconcentrated and the residue digested in hot EtOH (25 mL) to give asuspension, which was treated with water (25 mL) and left standing at23° C. for 2 h. The precipitate was then collected via filtration andwashed with water (20 mL). Drying the solid under reduced pressureprovided3′-bromo-4′-methyl-6′-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-4,5-dihydro-2H-spiro[furan-3,7′-pyrrolo[3,4-b]pyridine]-2′,5′(1′H,6′H)-dione(Compound 2078, 1.65 g, 3.690 mmol, 85.94% yield) as an off-white solid:ESMS (M+H) 447.17; ¹H NMR (DMSO-d₆, 300 MHz) δ 13.20 (br s, 1H), 8.14(s, 1H), 7.78 (s, 1H), 5.17 (q, J=9, 2H), 4.14 (d, J=9, 1H), 3.98 (m,2H), 3.83 (m, 1H), 2.59 (s, 3H), 2.49 (m, 1H), 2.32 (m, 1H).

As shown in step 24-vi of Scheme 24, to a suspension of Compound 2078(1.65 g, 3.690 mmol) in EtOH (50 mL) at 23° C. was added triethylamine(1.120 g, 1.543 mL, 11.07 mmol), followed by the addition of Pd/C (430mg, 0.4041 mmol) (10 wt % dry basis, Degussa type, wet). The reactionvessel was evacuated and the reaction atmosphere replaced with hydrogengas. After stirring for 16 hours, the reaction mixture was treated withMeOH and DCM (50 ml, each) and then filtered through diatomaceous earth,which was subsequently washed with 4:1 DCM:MeOH (100 mL). The combinedfiltrate was concentrated under reduced pressure to provide4-methyl-6-[1-(2,2,2-trifluoroethyl)pyrazol-4-yl]spiro[1H-pyrrolo[3,4-b]pyridine-7,3′-tetrahydrofuran]-2,5-dioneas a white solid (Compound 2079, 2 g, triethylamine impurity), which wasused as is in subsequent reactions: ESMS (M+H) 369.30; ¹H NMR (DMSO-d₆,300 MHz) δ 8.16 (s, 1H), 7.80 (s, 1H), 6.34 (br s, 1H), 5.17 (q, J=9, 2H), 4.13 (d, J=12, 1H), 3.93 (m, 3H), 2.45 (s, 3H), 2.30 (m, 2H).

As shown in step 24-vii of Scheme 24, To a solution/suspension ofCompound 2079 (1.359 g, 3.69 mmol, estimated mass of starting materialbased on 100% conversion in step 24-yl) in DCM (30 mL) was added DIEA(1.431 g, 1.929 mL, 11.07 mmol) followed by the addition ofN-(5-chloro-2-pyridyl)-1,1,1-trifluoro-N-(trifluoromethylsulfonyl)-methanesulfonamide(Commin's reagent, 1.594 g, 4.059 mmol). The reaction mixture becomeshomogenous in <10 minutes. After stirring for 3 h, additional Commin'sreagent (400 mg) was added and the reaction mixture stirred anadditional 90 minutes. The mixture was concentrated, loaded directlyonto a silica gel chromatography column in DCM (15 mL), and purified bymedium pressure silica gel chromatography (0-50% EtOAc in hexanes). Therecovered product was contaminated with Commin's reagent so it wasdissolved in DCM (100 mL), washed with saturated aqueous sodiumbicarbonate (100 mL), dried (magnesium sulfate), filtered, andconcentrated under reduced pressure to provide4′-methyl-5′-oxo-6′-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-4,5,5′,6′-tetrahydro-2H-spiro[furan-3,7′-pyrrolo[3,4-b]pyridine]-2′-yltrifluoromethanesulfonate (Compound 2080, 1.757 g, 3.436 mmol, 93.14%)as a yellow oil: ESMS (M+H) 501.24; ¹H NMR (DMSO-d₆, 300 MHz) δ 8.29 (s,1H), 7.90 (s, 1H), 7.64 (s, 1H), 5.22 (q, J=9, 2 H), 4.24 (d, J=12, 1H),4.07 (m, 1H), 3.90 (m, 2H), 2.74 (s, 3H), 2.39 (m, 2H).

As shown in step 24-viii of Scheme 24, a microwave reaction vessel wascharged with Compound 2080,3-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (121.4mg, 0.5164 mmol), sodium carbonate (136.8 mg, 1.291 mmol), DMF (5 mL),and water (2.5 mL). A stream of nitrogen was passed through the reactionmixture for 5 min, then Pd(PPh₃)₄ (24.87 mg, 0.02152 mmol) was added.Nitrogen gas was bubbled though mixture for a further 3 minutes and thenthe reaction vessel was sealed with a septum and heated to 105° C. (sandbath). After 16 hours at this temperature, the reaction mixture waspartitioned between EtOAc and water (100 mL each). The organics wereseparated, washed with brine (50 mL), dried (magnesium sulfate),filtered, and concentrated. The residue was purified by medium pressuresilica gel chromatography (0-8% EtOH in DCM) to yield a crude product,which was dissolved/suspended in hot EtOAc (2 mL), treated with hexanes(3 mL), and left standing at 23° C. for 30 minutes. The resultingprecipitate was collected by filtration and dried under reduced pressureto provide2-(5-methoxy-3-pyridyl)-4-methyl-641-(2,2,2-trifluoroethyl)pyrazol-4-yl]spiro[pyrrolo[3,4-b]pyridine-7,3′-tetrahydrofuran]-5-one(Compound 972, 87 mg, 0.1866 mmol, 43% yield) as an off-white solid:ESMS (M+H) 460.35; ¹H NMR (DMSO-d₆, 300 MHz) δ 8.93 (d, J=3, 1H), 8.36(d, J=3, 1H), 8.26 (s, 1H), 8.04 (s, 1H), 8.01 (dd, J=3, 3, 1H), 7.88(s, 1H), 5.16 (q, J=9, 2H), 4.19 (d, J=9, 1H), 4.05 (m, 3H), 3.88 (s,3H), 2.66 (s, 3H), 2.36 (m, 2H).

Using the appropriate intermediates, Compounds 978 and 989 were producedby a similar procedure.

Table 1 provides analytical characterization data for compounds offormula I (blank cells indicate that the test was not performed).

Lengthy table referenced here US20120040950A1-20120216-T00001 Pleaserefer to the end of the specification for access instructions.

Biological Assay of Compounds of the Invention Example 25 PI3KInhibition Assay

Using a Biomek FX from Beckman Coulter, 1.5 μL of each of ten 2.5-foldserial dilutions of a compound of the invention in 100% DMSO was addedto an individual well (hereafter, “test well”) in a 96 well polystyreneplate [Corning, Costar Item No. 3697]. One test well also contained 1.5μL of DMSO with no compound. Another well contained an inhibitor in DMSOat a concentration known to completely inhibit the enzyme, (hereafter“background well”). Using a Titertek Multidrop, 50 μL of Reaction Mix[100 mM HEPES pH 7.5, 50 mM NaCl, 10 mM DTT, 0.2 mg/mL BSA, 60 μMphosphatidylinositol(4,5)bisphosphate diC16 (PI(4,5)P₂; Avanti PolarLipids, Cat. No. 840046P) and PI3K isoform of interest (see Table 3 forisoform concentrations)] was added to each well. To initiate thereaction, 50 μL of ATP Mix [20 mM MgCl₂, 6 μM ATP (100 μCi/μmole³³P-ATP)] was added each well, followed by incubating the wells for 30min. at 25° C. Final concentrations in each well were 50 mM HEPES 7.5,10 mM MgCl₂, 25 mM NaCl, 5 mM DTT, 0.1 mg/mL BSA, 30 μM PI(4,5)P₂, 3 μMATP, and the PI3K isoform of interest (see Table 2). Final compoundconcentrations in each well ranged from 10 μM to 1 nM.

TABLE 2 PI3K Isoform Concentrations PI3K-α PI3K-β PI3K-γ PI3K-δ Enzymeconcentration in Reaction 4 nM 20 nM 4 nM 4 nM Mix Final enzymeconcentration 2 nM 10 nM 2 nM 2 nM

After incubation, the reactions in each well were quenched by additionof 50 μL of stop solution [30% TCA/Water, 10 mM ATP]. Each quenchedreaction mixture was then transferred to a 96 well glass fiber filterplate [Corning, Costar Item No. 3511]. The plate was vacuum-filtered andwashed three times with 150 μL of 5% TCA/water in a modified Bio-TekInstruments ELX-405 Auto Plate Washer. 50 μL of scintillation fluid wasadded to each well and the plate read on a Perkin-Elmer TopCount™ NXTliquid scintillation counter to obtain ³³P-counts representinginhibition values.

The value for the background well was subtracted from the value obtainedfor each test well and the data were fit to the competitive tightbinding Ki equation described by Morrison and Stone, Comments Mol. Cell.Biophys. 2: 347-368, 1985.

Each of the following compounds has a Ki of less than 0.1 micromolar forthe inhibition of PI3K-gamma: 1, 3, 6, 10, 12, 16, 18-20, 24-26, 34-36,38, 44, 46-48, 51-59, 64, 66-68, 70-73, 77-82, 85-109, 113-119, 122-124,126, 128-133, 135-136, 138-141, 143-156, 158, 161, 165-166, 170, 172,174-176, 178, 181-187, 194, 198-201, 204-219, 221-223, 225-227, 229-230,234, 236-237, 239, 241, 243-247, 250-273, 275-285, 287-312, 315, 317,320-322, 324-328, 330-332, 336-340, 342-347, 363, 365-366, 368-388,388-389, 391-407, and 409-411, 412-424, 426-437, 439-455, 457-461,463-464, 466, 468-472, 474-478, 480-481, 483-495, 497, 499-506, 508-573,576-584, 587-607, 609, 612, 614-616, 618-623, 625-658, 661, 664,666-672, 674-681, 683-685, 687-688, 690-695, 698-706, 708-710, 712-730,732-740, 742-749, 754-758, 760-762, 764-770, 772-779, 781-787, 789-801,802-803, 810-818, 822-825, 829-833, 835-838, 840, 843-871, 873, 875-893,896, 899-900, 902-906, 909-911, 913-916, 918-933, 935-943, 945-950,952-956, 958-984, 986-999.

Each of the following compounds has a Ki of 0.1 micromolar to 0.49micromolar for the inhibition of PI3K-gamma: 2, 5, 7, 9, 13-15, 17, 22,23, 27-33, 37, 39-43, 45, 49-50, 60-63, 65, 69, 74-76, 83, 110, 112,120, 125, 127, 134, 137, 142, 157, 159-160, 162, 164, 167-168, 171, 173,177, 179-180, 188-193, 196-197, 202-203, 224, 228, 231-232, 240, 242,248, 286, 313, 316, 318-319, 329, 333-335, 341, 348, 364, 367, 390, 408,425, 456, 462, 465, 467, 473, 479, 482, 496, 507, 574-575, 585-586, 608,611, 613, 617, 624, 660, 662, 686, 689, 696, 697, 707, 711, 731, 741,750, 771, 780, 802, 809, 819, 820-821, 834, 839, 872, 874, 894-895,897-898, 901, 907-908, 912, 934, 944, 951, and 957.

Each of the following compounds has a Ki of 0.5 micromolar to 2.5micromolar for the inhibition of PI3K-gamma: 4, 11, 21, 84, 111, 121,163, 169, 195, 220, 233, 235, 238, 249, 274, 314, 323, 610, 659, 663,665, 673, 682, 751, 759, 763, 826, 841, 842, 917, and 985.

Example 26 Microglia Activation Assay

Female C57B1/6J mice (7 weeks old) were purchased from JacksonLaboratory (Maine, US). Animals were acclimated for a week at standardlaboratory conditions (12 hrs light cycles) with free access to rodentchow and water. All procedures were in accordance with the NationalInstitute of Health Guidelines for the care and Use of LaboratoryAnimals and were approved by IACUCC. The endotoxin Lipopolysaccharide(LPS) (E. coli 011:B4, cat#437627) was purchased from Calbiochem. LPSwas dissolved in PBS buffer at a concentration of 0.05 mg/ml and storedin frozen aliquots. At the start of the study, mice received anintraperitoneal (i.p.) injection of LPS (0.5 mg/kg) for threeconsecutive days. Therapeutic treatment with VRT compounds was startedtogether with the 2^(nd) LPS administration and maintained throughoutthe study. Compound was dosed twice a day orally by gavage for a totalof 4 doses. 24 h following the last LPS injection, and 2 hrs followingthe last VRT dose, animals were sacrificed by CO₂ asphyxiation.

Following sacrifice, brains were rapidly removed and fixed overnight in10% neutral buffered formalin. Brains were then processed for routinehistology in an automated processor (Shandon Excelsior ES, ThermoScientific) and embedded in paraffin. IHC analysis was performed on 5 μmsections in the Ventana Benchmark System (Ventana Medical Systems Inc,Tucson, Ariz.) using prediluted antibodies to Iba1 (Wako chemical USA)at a dilution of 1:800. 3,3′-Diaminobenzidine (DAB) was used as achromogenic substrate, and the slides were counterstained withhaematoxylin.

Digital images were captured using the Aperio ScanScope Slide Scanner(Aperio Technologies, Vista, Calif.). Images were captured at 20×optical magnification, and analyzed using the software DefiniensDeveloper XD. Algorithms were created to count activated microglialcells taking into account distinct morphological characteristics ofactivated cells when compared to resting microglia. Compound efficacywas calculated as percent decrease in number of activated microgliarelative to vehicle control. For compound 271, a 39 percent decrease wasobserved at 10 mg/kg b.i.d. dosing. For compound 568 a range of 44 to 60percent decrease in the number of activated microglia at 10 mg/kg b.i.d.dosing was observed in three separate experiments. For compound 410 arange of 23 to 33 percent decrease in the number of activated microgliaat 5 mg/kg b.i.d. dosing was observed in three separate experiments.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it will be readily apparent to those of ordinary skill inthe art in light of the teachings of this invention that certain changesand modifications may be made thereto without departing from the spiritor scope of the appended claims.

LENGTHY TABLES The patent application contains a lengthy table section.A copy of the table is available in electronic form from the USPTO website(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20120040950A1).An electronic copy of the table will also be available from the USPTOupon request and payment of the fee set forth in 37 CFR 1.19(b)(3).

1. A compound having the formula:

or a pharmaceutically acceptable salt thereof, wherein: X¹ is N or CH;X² is N, CH, or C—CH₃; R¹ is selected from a phenyl ring, a 5-6 memberedheteroaryl ring, a pyridone ring, or a 9-10 membered fused bicyclicheteroaryl or heterocyclic ring system wherein each of said rings orring systems is optionally substituted with 1 or 2 independentoccurrences of R^(1a) and each of said heteroaryl or heterocyclic ringshas 1, 2, or 3 heteroatoms selected from nitrogen, oxygen, or sulfur;R^(1a) is chloro, fluoro, C₁₋₈aliphatic, —(CH₂)₀₋₂C₃₋₆cycloaliphatic,—(CH₂)₀₋₂-5-6 membered heterocyclic having up to two heteroatomsselected from nitrogen, oxygen, or sulfur, —CN, —C(O)C₁₋₄aliphatic,—C(O)NH(C₁₋₄aliphatic), —C(O)N(C₁₋₄aliphatic)₂, —C(O)OC₁₋₄aliphatic,—S(O)₂NH(C₁₋₄aliphatic), —S(O)₂N(C₁₋₄aliphatic)₂, or—S(O)₂C₁₋₄aliphatic, wherein up to 3 non-adjacent carbon atoms of saidaliphatic or cycloaliphatic of R^(1a) can be substituted for by —O—,—S—, or —N(R^(1b))— and wherein each of said aliphatic, cycloaliphatic,or heterocyclic of R^(1a) is optionally and independently substitutedwith up to 4 occurrences of J^(R); each J^(R) is independently fluoro,oxo, —(CH₂)₀₋₂CN, —(CH₂)₀₋₂CF₃, —C(O)R^(1b), —C(O)N(R^(1b))₂,—C(O)O(R^(1b)), —N(R^(1b))₂, —N(R^(1b))C(O)R^(1b), —(CH₂)₀₋₂OR^(1b),phenyl, or a 5-6 membered heteroaryl, 4-6 heterocyclyl, or 9-11 fusedbicyclic heteroaryl or heterocyclyl, each of said heteroaryl orheterocyclyl rings having up to 3 atoms selected from nitrogen, oxygen,or sulfur, wherein each of said cycloaliphatic, phenyl, heteroaryl, orheterocyclyl is optionally substituted with up to 2 R^(1c); each R^(1b)is, independently, selected from hydrogen, C₁₋₈aliphatic,—(CH₂)₀₋₁C₃₋₆cycloaliphatic, —(CH₂)₀₋₁C₄₋₆heterocyclic having up to twoheteroatoms selected from N or O, or two R^(1b) together with the atomto which they are bonded form a 5-6 membered heterocyclic ring, whereineach aliphatic, cycloaliphatic, or heterocyclic is optionallysubstituted with up to three F atoms or up to two —OH, —C₁₋₂alkyl, or—OC₁₋₂alkyl groups; each R^(1c) is, independently, fluoro, chloro,C₁₋₄aliphatic, —(CH₂)₀₋₂OH, —CN, —C(O)C₁₋₄aliphatic, or—C(O)OC₁₋₄aliphatic; R² is hydrogen, F, Cl, CF₃, C₁₋₂aliphatic,C₃₋₄cycloaliphatic, —N(CH₃)₂, —N(CH₂)₃, —OCF₃, —OCHF₂, or—OC₁₋₂aliphatic; R³ is hydrogen, C₁₋₆aliphatic, C₃₋₆ cycloaliphatic,C₄₋₇ heterocyclyl having 1 or 2 atoms selected from N or O,—(CH₂)₀₋₁CF₃, —OH, —OC₁₋₆aliphatic, —OC₃₋₆cycloaliphatic,—OC₃₋₆heterocyclyl having one oxygen atom, —O(CH₂)₂OC₁₋₂aliphatic, or—OC₁₋₂alkylC(O)OC₁₋₃aliphatic, or benzyl; and R⁴ is hydrogen orC₁₋₆alkyl; or R³ and R⁴ together with the carbon to which they arebonded form a 3-6 membered cycloaliphatic ring, a 3-6 memberedheterocyclic ring having up to two atoms selected from N or O, or aC₂alkenyl, wherein each of said aliphatic, cycloaliphatic, orheterocyclyl of R³, R⁴, or R³ and R⁴ together is optionally substitutedwith up to three F atoms, or up to two C₁₋₂alkyl, —C(O)C₁₋₄alkyl,—C(O)OC₁₋₄alkyl, —OH, or —OC₁₋₂alkyl groups; A is N or CR^(A); B is N orCR^(B), or A=B is a sulfur atom; C is N or CR^(C); D is N or CR^(D); Eis N or CR^(E) wherein no more than two of A, B, C, D, or E is N; R^(A)is hydrogen, CH₃, or OCH₃; R^(B) is hydrogen, F, Cl, C₁₋₃aliphatic,—(CH₂)₀₋₁CF₃, —(CH₂)₀₋₁CHF₂, or —O(CH₂)₀₋₁CF₃; R^(C) is hydrogen, F, Cl,C₁₋₃aliphatic, —(CH₂)₀₋₁CF₃, —(CH₂)₀₋₁CHF₂, N(R^(1b))₂, —OH,—O(CH₂)₀₋₁CF₃, or —OC₁₋₈aliphatic, wherein up to 2 non-adjacent carbonatoms of said aliphatic can be substituted for by —O—; R^(D) ishydrogen, fluoro, chloro, C₁₋₄aliphatic, —C(O)OH, —C(O)OC₁₋₄aliphatic,—C(O)N(R^(1b))₂, —CN, —C(R^(D1))═N—OR^(1b), —N(R^(1b))₂,—N(R^(D1))C(O)C₁₋₄aliphatic, —N(R^(D1))C(O)phenyl,—N(R^(D1))S(O)₂C₁₋₄aliphatic, —N(R^(D1))S(O)₂N(R^(1b))₂,—N(R^(D1))S(O)₂-phenyl —OH, —OC₁₋₈aliphatic,—O(CH₂)₀₋₁C₃₋₆cycloaliphatic, —SC₁₋₄aliphatic, —S(O)C₁₋₄aliphatic,—S(O)₂C₁₋₄aliphatic, or —S(O)₂N(R^(1b))₂; wherein up to 2 non-adjacentcarbon atoms of said aliphatic, cycloaliphatic, or heterocyclic of R^(D)can be substituted for by —O— and each of said aliphatic,cycloaliphatic, or phenyl of R^(D) can be substituted with up to 5fluorine atoms; or R^(D) and R^(C) together with the atoms to which theyare attached form a phenyl or pyridyl ring; each R^(D1) is,independently, hydrogen or C₁₋₂alkyl; and R^(E) is hydrogen, F, Cl,—NHC(O)C₁₋₈aliphatic, —OH, —OC₁₋₂aliphatic, —(CH₂)₀₋₁CF₃, —(CH₂)₀₋₁CHF₂,C₁₋₃aliphatic, C₃₋₄cycloaliphatic, N(R^(1b))₂, azetidin-1-yl. 2-7.(canceled)
 8. The compound according to claim 1 having the formula:

or a pharmaceutically acceptable salt thereof, wherein R¹is

wherein R^(1a) is —C₁₋₄alkyl, optionally and independently substitutedwith —CN, up to three F atoms, or up to two CH₃, —OC₁₋₂alkyl, or —OHgroups; R² is C₁₋₂alkyl; R³ is hydrogen, —OH, —OC₁₋₄ alkyl, or C₁₋₄alkyloptionally substituted with up to two —OH groups; R⁴ is hydrogen or CH₃,or R³ and R⁴ together form a C₃₋₆cycloalkyl ring optionally substitutedwith up to two OH groups, or a 4-6 membered heterocyclic ring having oneoxygen or anitrogen atom optionally substituted with C₁₋₄alkyl,—C(O)C₁₋₄alkyl, or C(O)O C₁₋₄alkyl; R^(C) is hydrogen, F, C₁₋₂alkyl, or—OC₁₋₂alkyl; and R^(D) is —OR^(D1), —C(O)N(R^(D1))R^(D2),—S(O)₂N(R^(D1))R^(D2), —S(O)₁₋₂R^(D2), —N(R^(D1))S(O)₂R^(D2), or—N(R^(D1))S(O)₂N(R^(D1))R^(D2), wherein R^(D1) is hydrogen or C₁₋₂alkyl,and R^(D2) is C₁₋₄alkyl, —(CH₂)₀₋₁C₃₋₆cycloalkyl, or—(CH₂)₀₋₁C₄₋₆heterocyclyl having up to two oxygen or nitrogen atoms,each alkyl, cycloalkyl, or heterocyclyl optionally substituted with upto three F atoms or up to two —OH groups.
 9. The compound according toclaim 8, or a pharmaceutically acceptable salt thereof, wherein R^(1a)is C₁₋₂alkyl, optionally substituted with up to 3 fluorine atoms. 10.The compound according to claim 8, or a pharmaceutically acceptable saltthereof, wherein R^(1a) is C₁₋₄alkyl, optionally substituted with CN.11. The compound according to claim 8, or a pharmaceutically acceptablesalt thereof, wherein R² is CH₃.
 12. The compound according to claim 8,or a pharmaceutical acceptable salt thereof, wherein at least one of R³and R⁴ is not hydrogen.
 13. The compound according to claim 12, or apharmaceutical acceptable salt thereof, wherein each of R³ and R⁴ isCH₃.
 14. The compound according to claim 12, or a pharmaceuticalacceptable salt thereof, wherein R³ and R⁴ together form a 4-6 memberedheterocyclic ring having one oxygen or a nitrogen atom optionallysubstituted with C₁₋₄alkyl, —C(O)C₁₋₄alkyl, or —C(O)OC₁₋₄alkyl. 15-19.(canceled)
 20. The compound according to claim 8, or a pharmaceuticallyacceptable salt thereof, wherein: R¹ is

R² is CH₃; R³ is hydrogen, C₁₋₂alkyl, OH, or OCH₃; R⁴ is hydrogen orCH₃; R^(C) is hydrogen; and R^(D) is —OC₁₋₂alkyl or —OC₃₋₅cycloalkyl,each optionally substituted with up to 3 fluorine atoms.
 21. Thecompound according to claim 20, or a pharmaceutically acceptable saltthereof, wherein R¹ is 1-(2,2-difluoroethyl)-1H-pyrazol-4-yl or1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl. 22-23. (canceled)
 24. Thecompound according to claim 23, or a pharmaceutically acceptable saltthereof, wherein: R¹ is

R² is CH₃; R³ is hydrogen, C₁₋₂alkyl, OH, or OCH₃; R⁴ is hydrogen orCH₃; R^(C) is hydrogen, F, Cl, C₁₋₃aliphatic, (CH₂)₀₋₁CF₃, —OCF₃, or—OC₁₋₈aliphatic; and R^(D) is —C(O)NHC₁₋₈aliphatic.
 25. (canceled) 26.The compound according to claim 8, or a pharmaceutically acceptable saltthereof, wherein each of R^(C) and R^(D) is —OCH₃.
 27. The compoundaccording to claim 8, or a pharmaceutical acceptable salt thereof,wherein R^(D) is —C(O)OH, —C(O)N(R^(1b))₂, —CN, —S(O)₂C₁₋₈aliphatic, or—S(O)₂N(R^(1b))₂.
 28. The compound according to claim 8, or apharmaceutically acceptable salt thereof, wherein each of R^(C) andR^(D) is, independently, hydrogen, fluoro, chloro, C₁₋₃aliphatic, CF₃,—OCF₃, —OCHF₂, or —OC₁₋₂aliphatic, wherein at least one of R^(C) andR^(D) is not hydrogen.
 29. The compound according to claim 28, or apharmaceutical acceptable salt thereof, wherein R^(C) is hydrogen andR^(D) is —OC₁₋₃alkyl, optionally substituted with up to 3 F atoms. 30.(canceled)
 31. The compound according to claim 1, or a pharmaceuticallyacceptable salt thereof, wherein said compound is a compound selectedfrom


32. A pharmaceutical composition comprising a compound according toclaim 1 and a pharmaceutically acceptable carrier, adjuvant, or vehicle.33-34. (canceled)
 35. A method of treating or lessening the severity ofa disease or condition selected from an autoimmune disease or aninflammatory disease of the brain or spinal cord selected from multiplesclerosis, epilepsy, Parkinson's Disease, Alzheimer's Disease,Huntington's Disease, or amyotrophic lateral sclerosis, comprising thestep of administering to said patient a compound or salt thereofaccording to claim 1, or a pharmaceutical composition thereof.
 36. Themethod according to claim 35, wherein said disease or disorder ismultiple sclerosis. 37-38. (canceled)
 39. A method of inhibitingPI3K-gamma kinase activity in a biological sample comprising contactingsaid biological sample with a compound according to claim 1, or acomposition comprising said compound.